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General Information
Symbol
Dmel\Ras85DV12.UAS
Species
D. melanogaster
Name
FlyBase ID
FBal0060587
Feature type
allele
Associated gene
Associated Insertion(s)
Carried in Construct
Also Known As
UAS-RasV12, RasV12, UAS-Ras85DV12, UAS-rasv12, UAS-Ras1V12, UAS RasV12, UAS-Dras1V12, Ras1V12, UAS-Ras85D.V12, Ras85DV12
Nature of the Allele
Mutations Mapped to the Genome
 
Type
Location
Additional Notes
References
Associated Sequence Data
DNA sequence
Protein sequence
 
 
Carried in construct
Cytology
Nature of the lesion
Statement
Reference
A constitutively active form of Ras85D is expressed under the control of UASt regulatory sequences.
Allele components
Product class / Tool use(s)
Encoded product / tool
Expression Data
Reporter Expression
Additional Information
Statement
Reference
 
Marker for
Reflects expression of
Reporter construct used in assay
Human Disease Associations
Disease Ontology (DO) Annotations
Models Based on Experimental Evidence ( 1 )
Modifiers Based on Experimental Evidence ( 1 )
Disease
Interaction
References
Comments on Models/Modifiers Based on Experimental Evidence ( 0 )
 
Phenotypic Data
Phenotypic Class
Phenotype Manifest In
dorsal mesothoracic disc & peripodial epithelium | somatic clone, with Scer\GAL4αTub84B.PL
eye disc & photoreceptor cell, with Scer\GAL4Act5C.PP
mesothoracic tergum & macrochaeta | ectopic, with Scer\GAL4sca-C253
mesothoracic tergum & macrochaeta | supernumerary, with Scer\GAL4sca-C253
muscle founder cell & visceral muscle primordium | ectopic, with Scer\GAL4twi.2PE
Detailed Description
Statement
Reference
Expressing Ras85DV12.UAS under the control of Scer\GAL4dpp.blk1 leads to wing disc overgrowths, with the widening of the Dpp expression domain and its cells showing an invasive phenotype.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Hml.Δ leads to a substantial increase in hemocyte count per larva, but pupae with this genotype exhibit similar eclosion rates at 25[o]C as compared to controls. None of these flies eclose when reared at 29[o]C, and pupae exhibit dessication and an absence of precursors of adult structures, in contrast to controls. Larvae expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Hml.Δ exhibit no significant difference in wound healing; larvae wounded and infected with the bacteria Staphylococcus aureus, Erwinia carotovora, or E.coli survive equally well as controls; and hemocytes isolated from larvae exhibit normal phagocytosis levels upon exposure to to heat killed E. coli, as compared to controls. Larvae expressing both Ras85DV12.Scer\UAS under the control of Scer\GAL4Bx-MS1096 or Scer\GAL4Hml.Δ infected with the nematode Heterorhabditis bacteriophora and its associated bacterium Photorhabdus luminescens, do not show a significant difference in survival as compared to controls; however, when under the control of Scer\GAL4Hml.Δ, infected (but not uninfected) larvae show a significant decrease in motility, as compared to controls.
Larvae expressing Ras85DV12.UAS under the control of Scer\GAL4Hml.Δ present extremely severe increase in circulating hemocytes and disrupted lymph gland primary lobes, but do not present defects in posterior signaling center size or morphology.
Whole eye-antennal disc clones that express Ras85DV12.UAS under the control of Scer\GAL4Act5C.PI form tumors that mostly stay in the CNS and rarely extend into the VNC.
Clones expressing Ras85DV12.Scer\UAS in third instar larval eye-antennal imaginal discs (under the control of Scer\GAL4Act.PU) or wing imaginal discs (under the control of Scer\GAL4tub.PU) do not lead to invasive tumors and do not lead to an increase in autophagosomes (assessed by a Atg8a fluorescence reporter) in either clone or neighboring disc tissue, as compared to controls; the eye-antennal imaginal disc clones do no exhibit an obvious increase in the proportion of cells entering cell cycle, and do not lead to an increase in autophagosomes in distant tissues - i.e. somatic muscles, midgut and fat body -, as compared to controls.
Expression of Ras85DV12.Scer\UAS driven by Scer\GAL4Scer\FRT.Act5C in clones induced in the eye-antennal discs leads to mild benign tumors in third instar larvae.
The expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI leads to a moderate overgrowth of the eye-antennal discs, without delaying pupal formation, as compared to controls.
Under control conditions, the clonal expression of Ras85DV12.UAS under the control of Scer\GAL4esg-NP7397 leads to a significant increase in the adult posterior midgut mitotic index in females and males, as compared to controls.
Expression of Ras85DV12.Scer\UAS driven by Scer\GAL4dpp.PU leads to tissue overgrowth in eye-antennal imaginal or wing discs; expression driven by Scer\GAL4Bx-MS1096 leads to tissue overgrowth in wing discs. Expression of Ras85DV12.Scer\UAS driven by Scer\GAL4Bx-MS1096 leads to changes in the mitotic cell cycle, with fewer cells in G1 or S phases and an increased fraction in the G2/M transition phase in third instar larval wing discs compared to wild type.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4esg-NP5130 dramatically increases the number of mitotic cells in the adult gut.
Adult midgut somatic clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4tub.PU have a significantly increased cell number as compared to control clones.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU does not affect the formation of cytoophidia (CTPsyn filaments) in follicle cells somatic clones in the egg chamber.
Animals expressing Ras85DV12.Scer\UAS via Scer\GAL4Hml.PG increases hemocyte count by approximately 100-fold.
Larval eye-antenna disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU overgrow to develop tumors. Cell death is detected in the wild type cells surrounding the clones. Eye disc clone cells expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU are able to undergo endocytosis, showing an increased level of dye internalisation compared with controls. The cells also show more efficient secretion: the rate of dye clearance is increased compared to controls. Generating clones expressing Ras85DV12.Scer\UAS in the whole eye disc (using the ey-FLP method) results in moderate overgrowth, compared to that seen in in mosaic eyes. The animals do not exhibit lethality.
Overexpression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Bx-MS1096 leads to an increase in the size of both wing imaginal discs and also the salivary glands in larvae and to pupal lethality. Dead pupae show signs of melanization focused around two areas in the dorsal part. Inhibition of Scer\GAL4 through the addition of Scer\GAL80ts.αTub84B rescues pupal lethality in a temperature-dependent manner. Larvae raised at 18[o]C pupate and eclose with wild-type wings while crosses shifted to 25[o]C give rise to flies with strongly melanized wings. Raising crosses at 29[o]C, where Scer\GAL80ts.αTub84B is inactive, re-establishes pupal lethality and the appearance of two melanotic spots in the anterior part of the wing. Overexpression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Bx-MS1096 induces Mmp1 expression and damage-associated signals (nuclear fragmentation, caspase activity). The integrity of the basement membrane in Ras85DV12.Scer\UAS expressing salivary glands are more severely affected than in imaginal discs from the same animal. Ras85DV12.Scer\UAS-expressing salivary glands exhibit an infiltration of haemocytes. However, the overall haemocyte count is not different from controls. Despite the presence of crystal cells attaching to the larval salivary glands, they do not exhibit any signs of melanization although this is subsequently observed in pupae. Ras85DV12.Scer\UAS-expressing salivary glands exhibit an increase in lamellocytes.
The expression of Ras85DV12.UAS under the control of Scer\GAL4Hml.Δ does not have an obvious effect on the growth, patterning and apoptosis levels (cleaved caspase 3 immunostaining) in the wing disc, which is in agreement with a normal adult wing, as compared to controls
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4phm.PO results in earlier pupation compared to controls. Most animals expressing Ras85DV12.Scer\UAS under the control of Scer\GAL45015 arrest at early pupal stages (P1-P9) with no detectable eye pigmentation or wings.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU at 21-22[o]C results in only approximately 40% of flies surviving to adulthood. Approximately 10% of the surviving adults have normal to nearly normal eye morphology, with the remaining adults having either hyperplastic or deformed eyes, or lacking one or both eyes. The number of mitotic cells per eye imaginal disc is increased compared to wild type in third instar larvae expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU.
Somatic clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU induced specifically in the eye disc show tumor overgrowth phenotype but the animals do not show any significant delay in pupariation.
Scer\GAL4C855a-mediated expression of Ras85DV12.Scer\UAS results in overgrown larval L3 neuroepithelium and ectopic neuroblasts. Scer\GAL4C855a-mediated expression of Ras85DV12.Scer\UAS in clones (via MARCM) produces cell-autonomous outgrowths and ectopic neuroblasts in the larval neuroepithelium.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4ap-md544 results in pupal lethality. More than 90% of mutants reach the prepupal stage.
Cells expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4en.PU show a significant increase in zippering speed during dorsal closure compared to neighbouring control cells. The number of filopodia at the leading edge is significantly increased in the mutant cells.
Fewer intestinal stem cell clones expressing Ras85DV12.Scer\UAS are seen per midgut over time. Clone cell polarity appears normal.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4byn-Gal4 (in combination with a Gal80[ts] transgene to restrict expression to the adult stage) for at least 7 days results in delamination of hindgut cells through the basal side of the epithelium, and evidence of basement membrane degradation, in contrast to controls. The delaminated hindgut cells progressively disseminate away from the hindgut to distant sites in the body, including the body wall, trachea, fat body and nephrocytes, increasing in penetrance and severity over time. This dissemination is not seen in the midgut when Ras85DV12.Scer\UAS is expressed under the control of Scer\GAL4esg-NP5130.
Expression of Ras85DV12.Scer\UAS in subperineurial glia under the control of Scer\GAL4Gli-rL82 increases the diameter of the peripheral nerves as compared with wild-type. Similarly to wild-type, plasmatocytes are not seen attached to peripheral nerves.
Mosaic clones overexpressing Ras85DV12.Scer\UAS in the gastric caeca of the midgut are enlarged and show reduced autophagy compared to surrounding wild type cells. Scer\GAL4Myo31DF-NP0001-mediated expression of Ras85DV12.Scer\UAS results in a large midgut with delayed degradation - gastric caeca still persist at +4 h relative puparium formation (RPF), and a less condensed midgut is observed at +12 h RPF compared to controls.
Expressing Ras85DV12.UAS under the control of Scer\GAL4ap-md544 results in a larger wing disc dorsal compartment in third instar larvae compared to wild-type controls.
Expression of Ras85DV12.Scer\UAS using Scer\GAL4Bx-MS1096 generates extra and ectopic veins in various wing regions.
Homozygous clones in the leg which express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU show autonomous activation of bract fate.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Mef2.PR results in an increase in cardioblast number in embryos compared to controls. Both blood progenitors and pericardial nephrocytes are also increased in number.
Expression of Ras85DV12.Scer\UAS in the midgut, under the control of Scer\GAL4esg-NP7397 results in the generation of many new midgut cells, including enterocyte-like cells. Expression of Ras85DV12.Scer\UAS in the midgut, under the control of Scer\GAL4Myo31DF-NP0001 and Scer\GAL80ts.αTub84B results in the generation of many new midgut cells, including enterocyte-like cells.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU in clones in the eye-antennal disc results in benign overgrowth of the clones in third instar larvae at 7 days after egg laying (AEL), but cells never invade into the nearby ventral nerve cord or other tissues. These animals can grow as larvae for up to 9 days AEL and then die as early pupae. Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU results in overgrowth of the eye.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4phm.PO results in severe larval growth defects, with shortened larval stages and small pupae. Scer\GAL4phm.PO->Ras85DV12.Scer\UAS prepupae preceed the appearance of control prepupae by up to 20 hours. These larvae also exhibit large and malformed prothoracic glands.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4wg-IS650 and Scer\GAL80ts.αTub84B activates EGFR signalling and blocks apoptosis in wing margin cells.
Expression of Ras85DV12.Scer\UAS in a MARCM third instar imaginal disc clone results in small tumours.
Clones of enterocytes overexpressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Myo31DF-NP0001 and Scer\GAL80ts.αTub84B rapidly disappear from the epithelium compared with wild-type clones suggesting an increased rate of delamination. The detached cells in these guts differ from those in wild-type flies in that they do not undergo cell death, have normal nuclei and do not contain enlarged vacuoles.
Clones of cells in the developing eye expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Scer\FRT.Act5C exhibit moderate benign overgrowth and never invade into the nearby ventral nerve cord or other tissues. Clones of cells in the developing eye expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Scer\FRT.Act5C in larvae bred on chloroquine or monensin exhibit invasive tumours. On day 18 AEL, 16.1% of chloroquine-treated tumours have prominently invaded into the ventral nerve cord, although the overgrowth is not notably enhanced. This invasive behavior is never observed in control tumours fed with solvent only medium. 11.8% of monensin-treated mutant animals develop ventral nerve cord invasion. Clones of cells expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Scer\FRT.Act5C display efficient lysosomal degradation.
Embryos expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4how-24B show an increase in the number of dorsal, dorsolateral and lateral adult muscle precursor cells compared to wild type. The number of ventral adult muscle precursor cells is unaffected in these embryos.
Eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU moderately overgrow. Marked overgrowth or invasion does not occur when the entire eye-antennal disc is composed of Ras85DV12.Scer\UAS-expressing cells. Inhibition of apoptosis in the overgrown eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU does not induce tumors. In contrast to wild type, where wounded wing discs are reduced in size compared to unwounded controls, wounded wing discs expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4nub.PK show a marked increase in overgrowth compared to the uninjured discs. No metastasis is detected.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4GMR.PF results in third instar larval eye discs with an enlarged lamina containing extra lamina cartridge neurons.
Expression of Ras85DV12.Scer\UAS in adult Malpighian tubule clones (using the MARCM system, under the control of Scer\GAL80αTub84B.PL and Scer\GAL4Scer\FRT.Act5C) results in renal and nephric stem cell over proliferation, and results in tumorous growth. The mitotic index of mutant clones is much greater than in wild-type clones. No cell death is detected the mutant clones. Mutant cells display disrupted polarity.
Ectopic expression of Ras85DV12.Scer\UAS driven by Scer\GAL4esg-NP7397 in adult midgut progenitor cells results in their overproliferation compared with control third instar larval midguts.
Scer\GAL4Orct2-cald-GAL4-mediated expression of Ras85DV12.Scer\UAS results in downregulation of autophagy and a strong persistent amnioserosa phenotype.
Expression of Ras85DV12.Scer\UAS within adult mushroom bodies (under the control of either Scer\GAL4c747 or Scer\GAL4c739) results in severe learning deficits.
Scer\GAL4btl.PS-mediated expression of Ras85DV12.Scer\UAS disrupts tracheal branching in the embryo.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4repo.PU induces 5-10 fold excess glia in the larval brain, and these glial are smaller than in wild type. Glial clones in adult brains expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4repo.PU contain two-fold more cells than wild type. Optic lobe clones expressing Ras85DV12.Scer\UAS generated using eyFLP under the control of Scer\GAL4repo.PU contain a modest number of excess and abnormal glia relative to wild type controls.
Expression of Ras85DV12.Scer\UAS in the larvae under the control of Scer\GAL4Act5C.Switch.PR results in lethality. Both male and female life-span are reduced by 80% when Ras85DV12.Scer\UAS is expressed in adult flies.
Expression of Ras85DV12.Scer\UAS in the wing blade using Scer\GAL4MD-638 causes pupal lethality, and the wings dissected from pharate adults are severely malformed and display excess vein differentiation.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PH early in the entire eye results in dramatic overgrowth and an increase in organ size.
Expression of Ras85DV12.Scer\UAS along the wing margin using Scer\GAL4vg.PU results in wing notching.
Primary cell cultures derived from embryos, and in which clones of cells that are expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP (and expressing an Avic\GFP marker) have been induced, show large clones of Avic\GFP-positive cells 10 days after clone induction (in contrast to control cultures where cells are induced to express the Avic\GFP marker alone). Most of the clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP are comprised of spindle-shaped cells and in 3-4 weeks, the cultures are confluent with Avic\GFP-positive cells expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP. The fraction of cells in S-phase is elevated and fewer cells die by apoptosis in primary cell cultures expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP compared to control cell cultures. Cells derived from embryos expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C give rise to a cell culture population that can be passaged for prolonged periods and appear immortalised and transformed.
The salivary glands of animals expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4fkh.PH are greatly increased in area compared to controls at 13.5 hours after puparium formation. At 24 hours APF, animals expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4fkh.PH still contain salivary gland tissue, in contrast to wild-type animals, where the salivary glands have completely degraded by this stage. Degradation is almost completely blocked in the mutant animals as intact salivary glands are still seen at 24 hours APF. The induction of autophagy that is seen in wild-type salivary glands at 13.5 hours APF is not seen in animals expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4fkh.PH. DNA fragmentation is detected by TUNEL in salivary glands of animals expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4fkh.PH at 13.5 hours after puparium formation, as occurs in control salivary glands at this stage.
Glia are normal in animals expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4GMR.PF. These animals have more plasmatocytes on the developing retina than wild type.
Clones in the eye antennal disc expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C do not show invasive behaviour.
The number of fusion competent myoblasts is decreased in stage 11 embryos that express Ras85DV12.Scer\UAS under the control of Scer\GAL4twi.PB.
Clones that express Ras85DV12.Scer\UAS behind the morphogenetic furrow of the eye disc, driven by Scer\GAL4GMR.PF, leads to disorganised clusters with many cells having upregulated adherens junction components.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Mhc.PW results in a microscopically normal larval muscle pattern, although these animals die as pupae.
Expression of Ras85DV12.Scer\UAS in somatic clones under the control of Scer\GAL4Act5C.PI results in activation of the JNK pathway and induction of benign tumours.
Expression of Ras85DV12.Scer\UAS, under the control of Scer\GAL4hs.2sev, causes a roughened eye phenotype with extra rhabdomeres in some ommatidia.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4dpp.3KK results in wing vein thickening.
Expression of Ras85DV12.Scer\UAS, under the control of Scer\GAL4en-e16E, in the posterior compartment cells of embryos leads to a reduction in the level of apoptosis of these cells, but has no effect on mitosis. Cells are of an increased size leading to an increase in compartment size of 29%.
Expression of Ras85DV12.Scer\UAS, under the control of Scer\GAL4da.G32, causes ectopic eye photoreceptor cell differentiation.
Clones of eye/antennal imaginal disc cells that express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI give rise to a mild tumor phenotype with no detectable invasiveness.
Ras85DV12.Scer\UAS Scer\GAL4αTub84B.PL somatic clones in the dorsal air sac primordium are larger than wild-type clones, forming bulging outgrowths. However, these clones never contribute to the primordium tip. (clones generated by Scer\GAL80 MARCM technique.)
Adults that express Ras85DV12.Scer\UAS under the control of Scer\GAL4amn-c651 are morphologically normal but are significantly reduced in size. A count of wing hair-cell number and wing hair-cell density shows that the reduced size phenotype is accompanied by a reduction in both cell number and cell size. Mutants that express Ras85DV12.Scer\UAS under the control of Scer\GAL4amn-X8 are very small but morphologically normal as third-instar larvae and pupae. However, such mutants fail to eclose. Larvae of this genotype develop more rapidly than normal and begin pupariation about 18 hours before wild-type larvae. Scer\GAL4amn-c651-driven Ras85DV12.Scer\UAS larvae begin wandering and pupariate about 24 hours before controls, although the pupariation time of these larvae is much more variable than in wild-type controls. Until about 84 hr AEL, these larvae are similar in size to wild-type larvae. At this time, about one-third of the Ras85DV12.Scer\UAS-expressing larvae appear to stop growing, begin pupariation, but fail to develop and ultimately die. The other two-thirds of these larvae grow, although more slowly than wild-type larvae, and form small pupae, which progress to become viable adults. Expression of Ras85DV12.Scer\UAS, under the control of Scer\GAL4amn-c651, increases prothoracic gland cell size, with no effect on cell number.
Expression of Ras85DV12.Scer\UAS, driven by Scer\GAL4sca-C253, leads to expression of ectopic bristles on the notum.
Cells in regions of the late third instar wing disc which are normally sensitive to irradiation induced apoptosis become resistant to it if they are part of Ras85DV12.Scer\UAS; Scer\GAL4Act5C.PI somatic clones. (40Gy of γ rays; assayed after 4hrs)
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4dpp.blk1 results in ectopic bristles and an excess of sensilla trichodea in the most proximal part of the coxa, ectopic bristles in the arista and third antennal segment, ectopic bristles in the capitellum and occasionally in the pedicellum of the haltere and ectopic thorn bristles in the female genitalia.
Stage 16 Ras85DV12.Scer\UAS; Scer\GAL4en-e16E embryos have more oenocytes per cluster than wild-type. Unlike rhoScer\UAS.cdCa; Scer\GAL4en-e16E the oenocyte clusters in these embryos do not show a multi-modal distribution of cell numbers with peaks corresponding to multiples of 3, suggesting that normal delamination cycles in groups of 3 are not occurring. In the resulting oenocyte clusters, terminal differentiation of some oenocytes is blocked or delayed.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4hs.PB using two 1 hour 37oC heat shocks at 14 hours after puparium formation (APF) and at 18 hours APF almost completely blocks the "late-stage" cell death which is normally seen in pupal retinas between 26-36 hours APF. However, "early-stage" cell death (which occurs between 18-24 hours APF) seems to be unaffected in these retinas.
4% of P{UAS-Ras85D.V12} males show rotated terminalia, rising to 55% when expression is driven by Scer\GAL4Pvf1-LP23.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4stop.Hml results in an elevation in the number of circulating larval hemocytes compared to wild type.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4sim.P3.7 in embryos does not result in a reduction in the number of midline glia.
When expression is driven by Scer\GAL4He.PZ, hemocyte proliferation is increased and lamellocyte and crystal cell numbers are unaffected. Melanotic masses form.
Late third instar larvae expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Cg.PA show a 40-fold increase in the number of hemocytes compared to wild type. These cells resemble plasmatocytes morphologically, but do not stain with a plasmatocyte-specific antigen. Cells that morphologically resemble lamellocytes are also seen in these larvae, and parasitisation of the larvae with L.boulardi results in induction of lamellocytes. Embryos carrying both Ras85DV12.Scer\UAS and Scer\GAL4Cg.PA do not have an increased number of hemocytes. 99% of animals expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Cg.PA die as early pupae under normal culture conditions at 25oC. However, 33% of these animals can survive to adulthood if cultured at very low densities. Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4en-e16E has no effect on the number and morphology of circulating hemocytes. When hemocytes expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Cg.PA are injected into the abdomens of wild-type females, the abdomens become enlarged and 64% of the injected flies die within 3 days. Hemocytes expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Cg.PA are capable of phagocytosing bacteria in an in vitro assay, although they are slightly less proficient than wild-type hemocytes.
Cells in Ras85DV12.Scer\UAS; Scer\GAL4αTub84B.PL somatic clones in the peripodial epithelium of the wing disc are transformed from squamous to columnar morphology. Gene expression in the clones indicate a transformation towards notum fate, rather than wing hinge or wing pouch.
When Ras85DV12.Scer\UAS is driven by Scer\GAL4HLHmδ ommatidial misrotations are seen in the eye.
Ras85DV12.Scer\UAS; Scer\GAL4twi.2PE embryos have increased numbers of visceral muscle founder cells.
Marked clones in the eye disc expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI are increased in size compared to control marked clones but are always located in the same areas as the control clones; mutant cells are not seen outside of the eye-antennal disc/optic lobe region, suggesting that tissue integrity is not compromised. The basement membrane is smooth and continuous on the outer surface of eye discs containing clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI.
Caspase activation in cells posterior to the furrow in eye discs is completely suppressed by Ras85DV12.Scer\UAS; Scer\GAL4GMR.PF. These discs also exhibit extensive ectopic photoreceptor differentiation posterior to the furrow.
Overexpression of Ras85DV12.Scer\UAS under the control of Scer\GAL4C380 causes a large increase in the number of synaptic boutons compared to overexpression of wild-type Ras85DScer\UAS.cKa.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP in wing disc cells results in an increase in cell size compared to controls. The proportion of cells in G1 is decreased compared to controls. Clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP are significantly more round than control clones.
Clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4αTub84B.PC in the wing disc (induced during the first larval instar) do not appear to stimulate growth or patterning of surrounding cells.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4GMR.PF in the eye disc results in all second mitotic wave cells dividing between columns 3-6 before a massive increase in photoreceptor differentiation.
When expression is driven by Scer\GAL4sca-C253, extra SMCs appear in proneural clusters and several extra macrochaetae are generated near extant ones.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4slbo.2.6 moderately affects posterior and dorsal border cell migration.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4dpp.blk1 results in ectopic furrow formation in the eye disc.
Imaginal disc cells expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP are larger than wild-type cells. The clones are round with smooth borders, mixing poorly with neighbouring wild-type cells. Elevated levels of cell death are not seen. The apparent cell doubling time is not affected. The proportion of cells in G1 is decreased and the proportion of cells in G2 and S phase is increased. 36 hours after induction, clones in imaginal discs expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP are 21% larger than control clones. Clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP (and control clones) induced during larval development are evenly distributed throughout developing vein and intervein regions of the wing approximately 24 hours after pupal formation. However, the nuclei of cells in the centre of some intervein clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP drop basally or disappear completely. Later in development (36 hours after puparium formation) most clones become concentrated in developing vein regions. When large clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP are induced in the wing, many intervein clones survive and form ectopic vein-like cuticle in adult wings.
When expression is driven by Scer\GAL4btl.PS in wild-type embryos, the primary branching pattern is not disrupted, however ectopic induction of downstream genes and extra terminal branching is seen.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4GMR.PF results in excessive neural determination of precursor cells in the eye disc.
Ubiquitously expressed Ras85DV12.Scer\UAS driven by Scer\GAL4e22c or Scer\GAL469B leads to the formation of extra denticles.
Clones expressing Ras85DV12.Scer\UAS in the eye disc under the control of Scer\GAL4Act5C.PP show ectopic photoreceptor formation anterior or posterior to the morphogenetic furrow.
Scer\GAL4dpp.blk1-mediated expression at 25oC causes lethality as early pupae, expression at 18oC causes lethality as young adults that exhibit severe developmental defects: mild rough eyes, severely malformed legs with missing or fused tarsal segments and smaller wings with broad L3 wing vein. Expression induces ectopic cell proliferation which leads to hyperplastic growth in imaginal discs that disrupts both the normal folding in the anterior compartment. Scer\GAL4dpp.blk1-mediated expression causes non-autonomous cell death.
When expressed under the influence of Scer\GAL4how-24B, midgut constriction 1 development is delayed or suppressed.
Embryos expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4en-e16E have ectopic denticles anterior to the denticle belts.
Scer\GAL4sli.PS-mediated expression causes ectopic midline glial cells.
Produces larvae with an excessive number of denticles, when expressed ubiquitously in embryos using Scer\GAL4arm.PS. Denticle rows 1-4 are normal, while small additional hooked denticles form anteriorly, and also posteriorly at the expense of denticles rows 5 and 6.
Lethal in combination with Scer\GAL4 lines expressed in the border cells. When expressed under the influence of Scer\GAL4hs.PB, delayed border cell migration is observed in 80% of stage 10B egg chambers. The time period for this effect is in stage 8 egg chambers. Little delay is seen when Ras85DV12.Scer\UAS is expressed in stage 9 egg chambers: border cells in the process of migrating are unaffected.
External Data
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Ras85DV12.UAS, Scer\GAL4hs.2sev has visible phenotype, enhanceable by S6kIID1/S6kII[+]
NOT Enhanced by
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Ras85DV12.UAS, Scer\GAL4GMR.PF, l(2)gl4 has large body phenotype, suppressible by Uev1A[+]/Uev1ADG14805
NOT suppressed by
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NOT Suppressor of
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Phenotype Manifest In
Enhanced by
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Ras85DV12.UAS, Scer\GAL4hs.2sev has ommatidium phenotype, enhanceable by S6kIID1/S6kII[+]
Ras85DV12.UAS, Scer\GAL4hs.2sev has rhabdomere phenotype, enhanceable by S6kIID1/S6kII[+]
NOT Enhanced by
Suppressed by
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Ras85DV12.UAS, Scer\GAL4GMR.PF, l(2)gl4 has larva phenotype, suppressible by Uev1A[+]/Uev1ADG14805
NOT suppressed by
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Ras85DV12.UAS, Scer\GAL4Mhc.PW is an enhancer of larval somatic muscle & cell phenotype of Hsap\FOXO1::Hsap\PAX7UAS.cGa, Scer\GAL4Mhc.PW
NOT Enhancer of
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NOT Suppressor of
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Ras85DV12.UAS, Scer\GAL4Act5C.PI/Scer\GAL4Act5C.PI, scrib1 has basal lamina & eye disc | somatic clone phenotype
Ras85DV12.UAS, Scer\GAL4Act5C.PI/Scer\GAL4Act5C.PI, scribunspecified has basal lamina & eye disc | somatic clone phenotype
Additional Comments
Genetic Interactions
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Reference
The co-expression of pnutGD1512 and Ras85DV12.UAS from the second instar larval stage under the combined control of Scer\GAL4ap-md544 and Gal80[ts] leads to the formation of tumors in the third instar larval wing disc.
Eye disc clones expressing Ras85DV12.UAS under the control of Scer\GAL4unspecified and are simultaneously homozygous for either Alg3tid2 or Alg9c02021 show non-invasive overgrowths, as compared to either single mutant clones.
Co-expression of l(2)glKK100777 or scribKK101128 exacerbates the increased hemocyte count observed in larvae expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Hml.Δ; these larvae are viable up to pupal stage and prepare for pupation, but only a subset of these pupae successfully eclose at 25[o]C, as compared to controls. None of these flies eclose when reared at 29[o]C, and pupae exhibit dessication and an absence of precursors of adult structures, in contrast to controls. Larvae expressing both Ras85DV12.Scer\UAS and l(2)glKK100777 or both Ras85DV12.Scer\UAS and scribKK101128 under the control of Scer\GAL4Hml.Δ exhibit no significant difference in wound healing; larvae wounded and infected with the bacteria Staphylococcus aureus, Erwinia carotovora, or E.coli survive equally well as controls; and hemocytes isolated from larvae exhibit normal phagocytosis levels upon exposure to to heat killed E. coli, as compared to controls. Larvae expressing both Ras85DV12.Scer\UAS and l(2)glKK100777 or both Ras85DV12.Scer\UAS and scribKK101128 under the control of Scer\GAL4Bx-MS1096 infected with the nematode Heterorhabditis bacteriophora and its associated bacterium Photorhabdus luminescens, do not show a significant difference in survival as compared to controls; but when these constructs are driven under the control of Scer\GAL4Hml.Δ, infected larvae show a significant increase in mortality as compared to controls, and also show a similar decrease in motility upon nematode infection as seen in those expressing only Ras85DV12.Scer\UAS.
Clones homozygous for scrib673 and expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU in third instar larval eye-antennal imaginal discs form invasive tumors, which display increased proportions of clone cells entering the cell cycle (but not undergoing mitosis), undergoing apoptosis and presenting mitochondrial defects - i.e., significant increases in the average mass and number of mitochondria per cell, a strong increase in the proportion of burst mitochondria, and a significant increase in the levels of reactive oxygen species, as compared to controls; the presence of these tumor clones also leads to a cell non-autonomous increase in autophagosomes (assessed by a Atg8a fluorescence reporter) in both neighboring disc tissue and distant tissues - i.e. somatic muscles, midgut and fat body - and in significant decreases in the respiratory capacity in both tumor and neighboring disc tissue, as compared to controls. scrib673-homozygous, Ras85DV12.Scer\UAS-expressing eye-antennal imaginal disc clone tumors are significantly suppressed by Atg13Δ81 homozygosity in clone cells (leading to reductions in the proportions of clone cells entering the cell cycle, but not undergoing mitosis or apoptosis, and leading to a reduction in the tumor size, but not invasiveness), in neighboring disc cells (leading to reductions in the tumor size and in the proportion of cell entering the cell cycle, but not undergoing mitosis or apoptosis), in the full organism except the clone cells (leading to reductions in the tumor size and invasiveness), or in the full organism (leading to reductions in the proportions of cells entering the cell cycle and undergoing mitosis, but not undergoing apoptosis, and leading to reductions in the tumor size and invasiveness); these tumors are also significantly suppressed by Atg14Δ5.2 homozygosity in the neighboring disc cells (leading to a reduction in the tumor size) or by the combination of Atg13Δ81 homozygosity in the tumor cells and Atg14Δ5.2 homozygosity in the neighboring disc cells. One copy of Atg13+tCH322-168O18 in the full organism suppresses the decrease in tumor size, but not invasiveness, resulting from Atg13Δ81 homozygosity in the full organism or in the full organism apart from the clone cells; one copy of Atg13ey.3.5 in the full organism also partially suppresses the decrease in tumor size resulting from Atg13Δ81 homozygosity. These tumors grow to significantly smaller sizes when grafted into the abdomen of either Atg14Δ5.2/Atg14EY14568 transheterozygous adult hosts (but not if the hosts possess one copy of Atg14+tCH322-175F03) or Atg8ad4 homozygous adult hosts, as compared to control hosts. The cell non-autonomous increase in autophagosomes resulting from the presence of clones homozygous for scrib673 and expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU in third instar larval eye-antennal imaginal discs is suppressed by Atg13Δ81 homozygosity in either the full organism (leading to suppression in distant tissues - somatic muscles, midgut and fat body) or in the full organism apart from the clone cells (leading to suppression in neighboring disc tissue), and suppressed by the clonal co-expression of either domeΔCYT.Scer\UAS (leading to suppression in neighboring disc tissue) or bskDN.Scer\UAS.cUa (leading to suppression in the midgut, but not in the somatic muscles or the fat body), but not suppressed by Stat92E85C9 homozygosity in the full organism apart from the clone cells. Clones homozygous for scrib673 and expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4tub.PU in third instar larval wing imaginal discs lead to a cell non-autonomous increase in autophagosomes (assessed by a Atg8a fluorescence reporter) in neighboring disc tissue, as compared to controls. Third instar larval eye-antennal imaginal disc clones homozygous for scrib1 and expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU form invasive tumors; the presence of these clones in egr1/egr3 transheterozygotes, or additional clonal homozygosity for kay3, or additional clonal co-expression of Ets21CKK103211 in combination with kaydsRNA.Scer\UAS.cUa do not lead to significant cell non-autonomous induction of autophagosomes (assessed by a Atg8a fluorescence reporter) in neighboring disc cells. Third instar larval eye-antennal imaginal disc clones co-expressing Ras85DV12.Scer\UAS and dlg1GD4689 under the control of Scer\GAL4Act5C.CoinFLP-FRT, in combination with Dicer-2 for efficient RNAi, form invasive tumors and induce autophagosomes in a cell non-autonomous manner in neighboring disc tissue (assessed by a Atg8a fluorescence reporter), as compared to controls; both the tumor formation and the cell non-autonomous autophagosome induction are suppressed by the additional clonal co-expression of either sdJF02514 or ykiHMS00041; the volume of these tumors is also significantly reduced by the clonal co-expression of either slifUY681 or Pi3K92ED954A.Scer\UAS.T:Hsap\MYC under the control of Scer\GAL4Act5C.CoinFLP-FRT; the cell non-autonomous increase in autophagosomes is also suppressed by the clonal co-expression of bskDN.Scer\UAS.cUa, but not of ImpL2dsRNA.Scer\UAS.cUa or Pi3K92ED954A.Scer\UAS.T:Hsap\MYC. The presence of third instar larval eye-antennal imaginal disc clones co-expressing Ras85DV12.Scer\UAS with either upd1[Scer\UAS.cUa] or upd3[d04951], but not ImpL2[s.Scer\UAS], under the control of Scer\GAL4Act.PU lead to a cell non-autonomous increase in autophagosomes (assessed by a Atg8a fluorescence reporter) in neighboring disc tissue, as compared to controls.
The comparatively weak tumor induction observed in clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Scer\FRT.Act5C induced in eye-antennal discs in third instar larvae is strongly enhanced when the clones are also homozygous mutant for fmt1. With continuous tumor progression almost half of the larvae show invasion of the tumor tissue into the ventral nerve cord at 11 days after egg laying (AEL). Number of mitotic cells is also highly increased compared to either of the single mutant clones, but no increase in the level of apoptosis is observed. The strong tumorigenic activity of Scer\GAL4Scer\FRT.Act5C>Ras85DV12.Scer\UAS;fmt1/fmt1 MARCM clones in third instar larval eye-antennal discs is strongly impeded and their invasive capacity abolished by co-expression of either bskDN.Scer\UAS or Tak1K46R.Scer\UAS in the clones. Similarly, the tumorigenicity of Ras85DV12.Scer\UAS-expressing clones is also enhanced if the clones are mutant for PpVΔ1, they display invasive behavior and increased mitotic activity; these characteristics are suppressed by co-expression of bskDN.Scer\UAS or Tak1K46R.Scer\UAS. Scer\GAL4Scer\FRT.Act5C>Ras85DV12.Scer\UAS;l(2)gl4/l(2)gl4 are also highly tumorigenic and have invasive capability and their overgrowth as well as invasiveness is significantly suppressed by co-expression of either fmtScer\UAS.cMa or PpVScer\UAS.P\T.N.T:Avic\GFP-EGFP and to even greater extent when both transgenes are expressed simultaneously in the mutant clones (however this effect is not due to increase in cell death). Co-expression of Ras85DV12.Scer\UAS and PpVGD7532 in somatic clones in the eye-antennal disc under the control of Scer\GAL4Act5C.PI results in mild tumorous overgrowth which is further enhanced by fmt1 heterozygosity.
The moderate overgrowth of eye-antennal discs bearing clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI is suppressed by Lkb1X5 homozygosity in the clones. The induction of larval eye-antennal disc clones both homozygous for Vps155 and expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI leads to an enhanced eye-antennal disc tumor phenotype, as compared to clones only expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI, and leads to delayed pupation, as compared to controls. Both these phenotypes are suppressed by additional Lkb1X5 homozygosity in the clones.
scrib1 in type II neuroblast lineage MARCM clones expressing Ras85DV12.Scer\UAS can be recovered at a similar rate as wild-type ones and instead of a single neuroblast they frequently contain 2 or 3 but the clones do not show significant overgrowth. scrib1 mutant clones in larval antennal discs clones expressing Ras85DV12.Scer\UAS (under the control of Scer\GAL4Dll-md23) all display tumor-like overgrowth.
The increased mitotic index in the adult posterior midgut induced by the clonal expression of Ras85DV12.UAS under the control of Scer\GAL4esg-NP7397 is not enhanced by clonal Apc2N175K, ApcQ8 double homozygosity alone or by clonal Apc2N175K, ApcQ8 double homozygosity in combination with clonal SxlKK116156 co-expression.
The high levels of apoptosis (detected by Caspase-3 staining) observed in Rbf15aΔ;Stam19 double homozygous somatic MARCM clones in third instar larval eye disc is strongly suppressed by expression of Ras85DV12.Scer\UAS under the control of the Scer\GAL4Act.PU driver in the mutant clones.
Co-expression of Dis3KK101473 significantly enhances tissue overgrowth in eye-antennal imaginal or wing discs with expression of Ras85DV12.Scer\UAS driven by Scer\GAL4dpp.PU or Scer\GAL4Bx-MS1096. Co-expression of sbrHMS02414 or CG10803KK104790 does not enhance tissue overgrowth in eye-antennal imaginal or wing discs with expression of Ras85DV12.Scer\UAS driven by Scer\GAL4dpp.PU or Scer\GAL4Bx-MS1096. Co-expression of Dis3KK101473 and Ras85DV12.Scer\UAS driven by Scer\GAL4Bx-MS1096 leads to mutual suppression of mitotic cell cycle changes in third instar larval wing discs with expression of either construct alone.
scrib1 eye discs expressing Ras85DV12.Scer\UAS in clones show a dramatic tumour-like overgrowth and metastasis. Eye discs co-expressing dlg1GD4689 and Ras85DV12.Scer\UAS in clones show tumour-like overgrowth and metastasis.
Transplanted scrib1 mutant eye disc fragments expressing MARCM closes of Ras85DV12.Scer\UAS grow continuously and induce distinctive bloating of the abdomen before killing the host. Wild-type discs grow only a limited amount before ceasing, and the transplanted host survives for weeks. Metastasis-like events are rare for the tumors. However, all the tumor-bearing hosts show robust wasting phenotypes in tissues distant from the transplant. Transplanted wild-type disc fragments do not induce these phenotypes. Cachexia-like wasting is observed in adipose tissue (fat body), muscle, and ovaries. Comparison of food consumption in wild-type and tumor-bearing hosts show that he tumor-induced wasting is not due to decreased food consumption (anorexia). Transplanted tissue fragments of Ras85DV12.Scer\UAS donors also expressing dlg1GD4689 under the control of Scer\GAL4ey.PU cause non-autonomous wasting. Expression of ImpL2GD6004 within the tumors of Scer\GAL4ey.PU>Ras85DV12.Scer\UAS,dlg1GD4689 origin significantly ameliorates the peripheral tissue wasting phenotypes. Hosts bearing dlg1GD4689, Ras85DV12.Scer\UAS, ImpL2GD6004 tumors show increased abdominal fat body mass. Muscle function assays further reveal improvements in both climbing ability and speed. There is a significant rescue of ovariole health, leading to restoration of egg production. The rescue is only observed when ImpL2GD6004 is expressed in the tumor. Transplanting dlg1GD4689, Ras85DV12.Scer\UAS tumors into ImpL2Def20 hosts does not ameliorate wasting.
l(2)gl4 mutant wing disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4tub.PU show increased levels of hypoxia compared to controls or either mutant alone. Ectopic tracheal structures are seen at the basal side of the disc, and in some cases several branches depart from a central mass and enclose a rudimentary lumen. l(2)gl4 Ras85DV12.Scer\UAS cells form mosaic tracheal branches with resident tracheal cells. Clone cells at the apical side of the disc are frequently interconnected with clone cells at the basal side by a bridge of tumour cells. When l(2)gl4 mutant clones expressing Ras85DV12.Scer\UAS are generated at 48 hours after egg laying (AEL) using a tracheal specific driver (Scer\GAL4btl.PU) no wing disc clones are observed. Tracheal clones are generated when clones are induced at 0-4 hours AEL but no overgrowth is observed.
Co-expression of Nhe2Scer\UAS.cGa and Ras85DV12.Scer\UAS under the control of Scer\GAL4ptc.PU results in a significant increase in the overall size of the wing disc compared with either transgene alone or wild type. There is also an increase in the number of invasive cells seen.
Larvae with eye-antennal disc clones co-expressing Ets21CKK103211 and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU exhibit developmental delay, a noticeable enlargement of clonal tissue that eventually outcompetes the non-clonal tissue, defective photoreceptor differentiation, and clonal tissue exhibits invasive behavior and infiltrates the brain lobes and the ventral nerve cord, as compared to controls. Clonal co-expression of bskDN.Scer\UAS.cUa suppresses the invasiveness of clonal cells, but does not suppress tumor formation or developmental delay seen with eye-antennal disc clones co-expressing Ets21CKK103211 and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. Larval eye-antennal disc clones co-expressing kayScer\UAS.cEa and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU exhibit hyperplasia but no invasive growth, as compared to controls, and pupae containing these clones die during the P4 or P5 stage. The majority of larvae with scrib1 mutant eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU exhibit defective photoreceptor differentiation, and developmental arrest as third instar giant larvae and die; only a few individuals form pseudopuparia and are developmentally delayed, starting on day 8 after egg laying. The cells of scrib1 mutant eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU are highly invasive, penetrating the ventral nerve cord of a majority of developmentally arrested larvae. Clonal co-expression of Ets21CKK103211 partially suppresses the lethality and developmental delay seen in larvae with scrib1 mutant eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU, but fails to suppress the invasive infiltration of the ventral nerve cord exhibited by cells of these clones. Clonal co-expression of ftz-f1JF02738 or kayN-Ala.Scer\UAS, or addition of a clonal kay3 mutation, but not co-expression of JradsRNA.Scer\UAS, partially suppresses the lethality and developmental delay seen in larvae with scrib1 mutant eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU, and suppresses the invasive infiltration of the ventral nerve cord exhibited by cells of these clones, although the tumor mass is not significantly decreased. Larvae with scrib1 mutant eye-antennal disc clones expressing ftz-f1JF02738, Ets21CKK103211, and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU reach pupation faster than larvae with scrib1 mutant eye-antennal disc clones expressing ftz-f1JF02738 and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU, or those with scrib1 mutant eye-antennal disc clones expressing Ets21CKK103211 and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. Clonal co-expression of kayN-Ala.Scer\UAS, or addition of a clonal kay3 mutation, but not co-expression of JradsRNA.Scer\UAS or ftz-f1JF02738, partially suppresses the defective photoreceptor differentiation seen in scrib1 mutant eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. Larvae with scrib1, kay3 mutant eye-antennal disc clones expressing Ras85DV12.Scer\UAS and Ets21CKK103211 under the control of Scer\GAL4Act.PU exhibit suppression of larval lethality as compared to either larvae with scrib1, kay3 mutant eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU, or those with scrib1 mutant eye-antennal disc clones expressing Ras85DV12.Scer\UAS and Ets21CKK103211 under the control of Scer\GAL4Act.PU; but these larvae still exhibit pupal lethality. Adults with scrib1 mutant eye-antennal disc clones expressing Ras85DV12.Scer\UAS and ftz-f1JF02738 under the control of Scer\GAL4Act.PU exhibit a rough eye that is increased in size, and contains fewer clonally-derived ommatidia, as compared to controls. Larval eye-antennal disc clones co-expressing ftz-f1Scer\UAS.β.T:Zzzz\FLAG and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU exhibit hyperplasia but no invasive growth, as compared to controls, and pupae containing these clones die during the P4 or P5 stage. Larval eye-antennal disc clones co-expressing ftz-f1Scer\UAS.α.T:Zzzz\FLAG and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU exhibit hyperplasia but no invasive growth, as compared to controls, and pupae containing these clones die during the P4 or P5 stage.
Expression of Ras85DV12.Scer\UAS suppresses the photoreceptor differentiation defects seen in Vps43B1 mutant clones. The increase in cell death seen in Vps43B1 mutant clones is also suppressed by Ras85DV12.Scer\UAS.
Expression of Ras85DV12.Scer\UAS suppresses the wing vein loss seen when PtpmegScer\UAS.cLa is expressed under the control of Scer\GAL4en.PU.
The increased cell proliferation observed upon expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4esg-NP5130 in the adult gut is suppressed by co-expression of either Sox21aJF02191 or kaydsRNA.Scer\UAS.
The presence of bun200B/bun200B suppresses the increased cell number seen in midgut clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4tub.PU.
Co-expression of Ras85DV12.Scer\UAS partially suppresses the pupariation delay and developmental arrest at the larval stage seen in Scer\GAL4phm.PO>Itp-r83AGD1676 (along with UAS-Dcr-2).
Co-expression of rlGD4697, gfzfGD10273, Prp19KK101335, Prp19GD11681, CG11266NIG.11266R or CG1603GD9160 reduces hemocyte proliferation induced by expression of Ras85DV12.Scer\UAS via Scer\GAL4Hml.PG.
Co-expression of Ras85DV12.Scer\UAS fails to rescue the increased apoptosis observed in clones expressing Rab5GD10492 under the control of Scer\GAL4hh.PU in the posterior compartment of the wing disc, however if there are many of these clones in the wing disc, they are able to form invasive tumors despite containing many cells in apoptosis.
Sec151 dramatically suppresses the overgrowth phenotype and pupal lethality seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. The increased secretion (dye clearance) seen in cells expressing Ras85DV12.Scer\UAS is also suppressed by Sec151. Cell death is no longer detected in the wild type cells surrounding the clones. Larval eye-antenna disc clones expressing Ras85DV12.Scer\UAS in a scribunspecified mutant background overgrow and invade the ventral nerve cord, killing the animal during the larval stages. Expression of Sec15KK101708 suppresses the overgrowth and ventral nerve cord invasion seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU in a scribunspecified mutant background. Expression of Sec15GD12109 suppresses the overgrowth and ventral nerve cord invasion seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU in a scribunspecified mutant background. Expression of Sec8KK101531 suppresses the overgrowth phenotype and pupal lethality seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. Expression of Sec6GD11616 suppresses the overgrowth phenotype and pupal lethality seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. Expression of bskDN.Scer\UAS suppresses the increased cell death seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU in a Sec151 mutant background. Tumor growth is not restored to that seen in clones expressing Ras85DV12.Scer\UAS alone. Expression of egrdsRNA.Scer\UAS suppresses the overgrowth seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. A egr1 mutant background suppresses the overgrowth seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. Expression of TaceGD572 suppresses the overgrowth seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. The phenotype is further suppressed when the adjacent cells are homozygous mutant for Tak11. Expression of TacedsRNA.Scer\UAS suppresses the overgrowth seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. The phenotype is further suppressed when the adjacent cells are homozygous mutant for Tak11. Expression of domeJF01682 suppresses the overgrowth seen in eye-antennal disc clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. The Ras85DV12.Scer\UAS, domeJF01682 clones have reduced mitotic potential compared to Ras85DV12.Scer\UAS alone. The phenotype is less severe when the clones are generated in the entire eye disc than when the clones are surrounded by wild type cells in mosaic clones, and yields viable adults. The adult flies co-expressing domeJF01682 have a smaller eye size than those expressing Ras85DV12.Scer\UAS alone.
Rare and extremely small homozygous ecdl(3)23 clones can be recovered if cell growth is enhanced in the clones by expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI.
A santa-maria1 homozygous mutant background partially suppresses the Ras85DV12.Scer\UAS-induced overgrowth, including the aberrant histology of the salivary glands and the mortality. Heat-shock expression of santa-mariahs.PW in flies expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Bx-MS1096 at 29[o]C results in melanization of the salivary glands in approximately 4% of cases, in contrast to larvae that express Ras85DV12.Scer\UAS alone at the same temperature and which do not show any signs of melanization.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Scer\FRT.Act5C in l(2)gl4 mutant cells in eye-antennal discs using the ey-FLP/MARCM system results in tumor-like growth with invasive migration into the ventral nerve cord of the CNS. The growth and invasion of these tumors is dramatically suppressed upon co-expression of pucScer\UAS.cMa. Expression of benGD1387 significantly suppresses the growth and invasion of the tumors seen when Ras85DV12.Scer\UAS is expressed in l(2)gl4 mutant cells in eye-antennal discs using the ey-FLP/MARCM system. One copy of ben1 partially suppresses the growth and invasion of the tumors seen when Ras85DV12.Scer\UAS is expressed in l(2)gl4 mutant cells in eye-antennal discs using the ey-FLP/MARCM system.
The apoptosis observed in dlg1A40.2 mutant wing disc tumors is virtually abrogated by the co-expression of egrKK103432 and Ras85DV12.UAS, but not by the expression of Ras85DV12.UAS alone, under the control of Scer\GAL4Hml.Δ.
Clones generated in the larval eye discs expressing Cskunspecified and Ras85DV12.Scer\UAS show limited tissue overgrowth. Expression of Nek2Scer\UAS.T:Avic\GFP enhances the tissue overgrowth seen in larval eye disc clones expressing Cskunspecified and Ras85DV12.Scer\UAS, and leads to the production of secondary tumours in the body of the larvae. This phenotype can be suppressed by feeding flies with the Akt1 inhibitor MZ2206; significantly smaller primary tumours are seen and there is a complete absence of secondary tumours. Injection of dissociated cells from Nek2Scer\UAS.T:Avic\GFP Cskunspecified Ras85DV12.Scer\UAS mutant eye discs (generated using ey-FLP MARCM) into the dorsal notum region of wild type adult flies results in detection of T:Avic\GFP positive cells in various parts of the adult body. Dispersed Avic\GFP-positive cells are not seen when flies are injected with eye disc cells that express Nek2Scer\UAS.T:Avic\GFP alone.
Co-expression of cncdsRNA.C.Scer\UAS in animals expressing Ras85DV12.Scer\UAS under the control of Scer\GAL45015 allows the majority of animals to develop to late pupal stages, and some eclose as adults. Co-expression of cncdsRNA.C.Scer\UAS suppresses the small body size of pupae expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4phm.PO.
Co-expression of BHDScer\UAS.T:Ivir\HA1 has no effect on the semi-lethality caused by expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU at 21-22[o]C. However, the eye overgrowth and deformation phenotypes seen in adult survivors is partially suppressed. Co-expression of BHDScer\UAS.T:Ivir\HA1 almost completely suppresses the increase in the number of mitotic cells per eye imaginal disc seen in larvae expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU. Co-expression of either BHDdsRNA.Sym.Scer\UAS or BHDGD6590 enhances the semi-lethality caused by expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU at 21-22[o]C, such that few adult escapers emerge. The eye defects of adult escapers are enhanced. Co-expression of either BHDdsRNA.Sym.Scer\UAS or BHDGD6590 exacerbates the increase in the number of mitotic cells per eye imaginal disc seen in larvae expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU. Rpt4G0345/+ partially suppresses the semi-lethality caused by expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU at 21-22[o]C. Co-expression of Rpt4Scer\UAS.cGa enhances the semi-lethality caused by expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU at 21-22[o]C. RpI135k16513/+ partially suppresses the semi-lethality caused by expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU at 21-22[o]C.
The tumor overgrowth of somatic clones (induced specifically in the eye disc) expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU is suppressed when the clones are also homozygous for either CskQ156Stop or Cskj1D8. Ras85DV12.Scer\UAS expressing CskQ156Stop mutant clones show highly increased levels of apoptosis, have an intact basement membrane and only very rarely form secondary tumors distant from the eye field in third instar larvae. The larvae display a delay in pupariation. The increased cell-death phenotype of CskQ156Stop/CskQ156Stop somatic clones (induced specifically in the eye disc) expressing Ras85DV12.Scer\UAS under Scer\GAL4Act.PU is fully suppressed by co-expression of InRA1325D.Scer\UAS, resulting in tumor overgrowth of the eye disc in nearly all larvae. Majority of the larvae fail to pupariate, the ones that manage do so with a significant delay. Somatic clones (induced specifically in the eye disc) expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU and homozygous mutant for scrib1 show substantial tumorigenic overgrowth but do not form secondary tumors although floating clone cells are frequently observed in the hemolymph.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4tub.PU in l(2)gl4 mutant wing disc mutant clones results in overproliferation and neoplastic transformation in both the proximal and distal domains.
Ras85DV12.Scer\UAS scrib1 tumour cells fail to differentiate into neurons. Loss of cher markedly interferes with the growth of Ras85DV12.Scer\UAS scrib1 tumours. The loss of cher does not affect the level of apoptosis in either the clonal nor the surrounding eye-antennal imaginal disc, indicating that loss of cher does not affect tumour cell viability. Eye-antennal imaginal discs carrying Ras85DV12.Scer\UAS scrib1 cher1 clones display extra cell divisions, mainly occurring in the surrounding non-clonal tissue. Despite sizeable Ras85DV12.Scer\UAS scrib1 cher1 clones clones located posterior to the morphogenetic furrow, the number of elav-positive differentiating photoreceptors increases and their organization is better than in Ras85DV12.Scer\UAS scrib1 clones. Consequently, well-ordered arrays of ommatidia appear in Ras85DV12.Scer\UAS scrib1 cher1 eye-antennal imaginal discs compared with the indiscernible pattern in Ras85DV12.Scer\UAS scrib1 discs. Many apically localised Ras85DV12.Scer\UAS scrib1 cells are elongated. Cross-sections of Ras85DV12.Scer\UAS scrib1 cher1 eye-antennal discs reveals that large tumours only reside within the basal compartment of the columnar epithelium. Smaller Ras85DV12.Scer\UAS scrib1 cher1 clones are confined between the squamous peripodial epithelium and the columnar epithelium. In contrast to highly invasive Ras85DV12.Scer\UAS scrib1 clonal tumours, fewer Ras85DV12.Scer\UAS scrib1 cher1 clones invaded the brain and the ventral nerve cord. Expression of cherKK107518 suppresses the tumour invasiveness found in Ras85DV12.Scer\UAS scrib1 clones. Larvae bearing Ras85DV12.Scer\UAS scrib1 tumours all die in the third instar and never pupate. In contrast, on day 8-9 after egg laying, 30% of Ras85DV12.Scer\UAS scrib1 cher1 larvae begin to wander and pupate, albeit 2-3 days later than control animals. Thirty percent of Ras85DV12.Scer\UAS scrib1 cher1 pupape emerge as adults. These adult flies exhibit abnormally enlarged eyes with a folded surface, accommodating surplus ommatidia. Expression of zipGD1566 lessens the invasiveness of Ras85DV12.Scer\UAS scrib1 tumours and improves pupation rate.
Expression of Ras85DV12.Scer\UAS in l(2)gl4 mutant cells in eye-antenna discs using the ey-FLP/MARCM system induces strong tumour-like growths, with invasive migration into the VNC 8 days after egg laying. Such animals keep growing as oversized larvae carrying huge tumours in their heads and die before pupation. Blocking JNK signaling by expression of pucScer\UAS.cMa dramatically suppresses the growth and invasion behaviours of Ras85DV12.Scer\UAS/l(2)gl4 tumours. Loss of Src42A, through expression of Src42ANIG.7873R, dramatically suppresses tumour cell invasion into the ventral nerve cord found in Ras85DV12.Scer\UAS/l(2)gl4 mutants and enables the animals to survive to the pupal stage, whereas the tumour size remains largely unaffected. Loss of Src64B, through expression of Src64BGD12263, dramatically suppresses tumour cell invasion into the ventral nerve cord in Ras85DV12.Scer\UAS/l(2)gl4 mutants and enables the animals to survive to the pupal stage, whereas the tumour size remains largely unaffected.
Expression of Ras85DV12.Scer\UAS in l(2)gl4 mutant developing eye-antennal discs, under the control of Scer\GAL4GMR.PF, results in tumour-like overgrowths, with invasive migration into the ventral nerve cord of the central nervous system. The mutant animals keep growing as oversized larvae with huge tumours in the head region and die before pupation. Inactivation of JNK signaling, through the expression of pucScer\UAS.cMa, dramatically suppresses the Ras85DV12.Scer\UAS/l(2)gl4 mutant tumour growth and invasion, and rescues animals to pupal stage. Expression of Uev1AGD6650 mutant background dramatically suppresses the Ras85DV12.Scer\UAS/l(2)gl4 mutant tumour growth and invasion, and rescues animals to pupal stage. A Uev1ADG14805 heterozygous mutant background dramatically suppresses the Ras85DV12.Scer\UAS/l(2)gl4 mutant tumour growth and invasion, and rescues animals to pupal stage.
Expression of taydsRNA.Scer\UAS enhances the ectopic wing vein phenotype seen when Ras85DV12.Scer\UAS is expressed under the control of Scer\GAL4salm.EPv. Expression of tayEP-866 suppresses the ectopic wing vein phenotype seen when Ras85DV12.Scer\UAS is expressed under the control of Scer\GAL4salm.EPv. Co-expression of Ras85DV12.Scer\UAS and rlSem.Scer\UAS.T:Ivir\HA1 under the control of Scer\GAL4salm.EPv results in a strong ectopic wing vein phenotype. Expression of tayFL.Scer\UAS.T:Zzzz\FLAG does not suppress the ectopic wing vein phenotype seen when Ras85DV12.Scer\UAS and rlSem.Scer\UAS.T:Ivir\HA1 are co-expressed under the control of Scer\GAL4salm.EPv.
Expression of PRL-1Scer\UAS.cPa enhances the lethality seen when Ras85DV12.Scer\UAS is expressed in the dorsal compartment of the wing disc under the control of Scer\GAL4ap-md544. Fewer than 10% of mutants reach the prepupal stage, compared to 90% when Ras85DV12.Scer\UAS is expressed alone. The larvae appear lethargic.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4crq.PO partially rescues the embryonic lethality of Df(2L)Pvf2-3/Df(2L)Pvf2-3 mutants.
Flies co-expressing Ras85DV12.Scer\UAS produce fewer intestinal stem cell (ISC) clones per midgut over time compared with Apc2g10 ApcQ8 mutants alone, similar to what is seen when Ras85DV12.Scer\UAS is expressed alone. Approximately 35% of the clones that remain undergo rapid proliferation and develop into large spherical-tumor cell masses (termed 'transformed clones'). There is no obvious increase in apoptosis in these transformed clones and the majority are found in the anterior or posterior midgut rather than the middle midgut. Few enterocytes (ECs) or enteroendocrine cells (ees) are seen, indicating that differentiation is blocked. The remaining 65% of clones are smaller than their wild type counterparts, but the ratio of ISCs to differentiated EC and ee cells is comparable to wild type. Approximately 10% of Apc2g10 ApcQ8, Ras85DV12.Scer\UAS mutant ISC clones extrude basally toward the surrounding muscle layer, although the underlying base layer remains unbroken. Expression of Ras85DV12.Scer\UAS induces polarity changes in Apc2g10 ApcQ8 mutant clones.
Ras85DV12.Scer\UAS-scrib1 mutant larval MARCM clones exhibit tissue overgrowth. Exposure to acivicin restrains Ras85DV12.Scer\UAS-scrib1 tumor overgrowth. Knockdown of CTPsyn, through expression of CTPsynGD4740, induces a strong, significant reduction in tumor growth in Ras85DV12.Scer\UAS-scrib1 mutant larval MARCM clones. Knockdown of CG9674, through expression of CG9674GD14285, induces a strong, significant reduction in tumor growth in Ras85DV12.Scer\UAS-scrib1 mutant larval MARCM clones. Knockdown of bur, through expression of burGD13797, induces a reduction in tumor growth in Ras85DV12.Scer\UAS-scrib1 mutant larval MARCM clones. Knockdown of r, through expression of rGD9607, induces a reduction in tumor growth in Ras85DV12.Scer\UAS-scrib1 mutant larval MARCM clones. Knockdown of ade2, through expression of ade2GD1356, induces a reduction in tumor growth in Ras85DV12.Scer\UAS-scrib1 mutant larval MARCM clones. Knockdown of Gfat1, through expression of Gfat1GD7732, induces a reduction in tumor growth in Ras85DV12.Scer\UAS-scrib1 mutant larval MARCM clones. Knockdown of Prat, through expression of PratGD9838, induces a reduction in tumor growth in Ras85DV12.Scer\UAS-scrib1 mutant larval MARCM clones. Knockdown of Prat2, through expression of Prat2GD16167, induces a reduction in tumor growth in Ras85DV12.Scer\UAS-scrib1 mutant larval MARCM clones. Knockdown of CG9940, through expression of CG9940GD14730, induces a reduction in tumor growth in Ras85DV12.Scer\UAS-scrib1 mutant larval MARCM clones.
Co-expression of bskDN.Scer\UAS, bskHMS00777, TimpScer\UAS.cPa or JraNIG.2275R partially suppresses the dissemination of hindgut cells seen in flies expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4byn-Gal4. Co-expression of hepAct.Scer\UAS or imdScer\UAS.cGa enhances the dissemination of hindgut cells seen in flies expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4byn-Gal4.
Larvae expressing Scer\GAL4Gli-rL82>Ras85DV12.Scer\UAS display nerves that are significantly wider than either Scer\GAL4Gli-rL82>Ras85DV12.Scer\UAS or egh62d18 single mutants.
Scer\GAL4Bx-MS1096-mediated expression of both Ras85DV12.Scer\UAS and PlzfScer\UAS.cMa generates significantly enhanced extra and ectopic veins as compared to expression of either transgene alone. This effect is more dramatic at 18[o]C than 25[o]C. The Scer\GAL4Bx-MS1096, Ras85DV12.Scer\UAS extra wing vein phenotype is partially suppressed in PlzfΔ145/+ or PlzfΔ11/+ heterozygotes.
Co-expression of Ras85DV12.UAS and Mkp3UAS.cUa in the midgut, under the control of Scer\GAL4NP7397 fails to induce midgut cell proliferation. Induction of activated Ras85D in Ras85DV12.UAS Stat92E397 MARCM clones drives the growth of large intestinal stem cells, regardless of the Stat92E mutant background.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU in clones in the eye-antennal disc (where the clones are also homozygous for sds22Δ2.8) results in extensive hyperproliferation of the clones in the eye-antennal disc at 7 days after egg laying (AEL), but in addition, clonal cells are also seen in the ventral nerve cord at low frequency. At 15 days AEL, significant numbers of clonal cells are seen spreading from a primary tumour in the brain into the ventral nerve cord. In addition, as the tumours grow, the two eye-antennal discs appear to fuse into one large mass. These animals can grow as larvae for up to 15 days AEL and die before pupation or as early pupae. Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU in clones in the eye-antennal disc (where the clones are also homozygous for scrib673) results in strong tumour growth at 7 days AEL. These animals grow as larvae until 13 days AEL and die before pupation. Co-expression of sds22Scer\UAS.cJa in these clones strongly suppresses the tumour growth phenotype in all clones observed at 7 days AEL and most of these animals can pupate, but die as early pupae. Eye overgrowth caused by expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4ey.PU is not suppressed by co-expression of sds22Scer\UAS.cJa.
Co-expression of Ras85DV12.Scer\UAS and Hr4Scer\UAS.cQa under the control of Scer\GAL4P0206 results in wild-type timing of pupariation, and a partial suppression of the Hr4Scer\UAS.cQa-mediated lethality. Hr4Scer\UAS.cQa co-expression with Ras85DV12.Scer\UAS represses the hyper-proliferation of the ring gland seen in Ras85D overexpression flies.
Co-expression of Pi3K92EGD11228 suppresses tumour growth in Ras85DV12.Scer\UAS MARCM third instar imaginal disc clones, although this suppression is weaker than that seen upon expression of Pi3K92EGD11228 in Ras85DV12.Scer\UAS-dlg1GD4689 MARCM third instar imaginal disc clones. Overexpression of ykiScer\UAS.cHa in Ras85DV12.Scer\UAS MARCM clones generates larger tumorigenic overgrowths than in either single mutant clone. Overexpression of upd1Scer\UAS.cZa in Ras85DV12.Scer\UAS MARCM clones generates larger tumorigenic overgrowths than in either single mutant clone. Expression of Ras85DV12.Scer\UAS and dlg1GD4689 in MARCM third instar imaginal disc clones results in a neoplastic phenotype. Co-expression of Pi3K92EGD11228 suppresses tumour growth in dlg1GD4689 MARCM third instar imaginal disc clones, although this suppression is weaker than that seen upon expression of Pi3K92EGD11228 in Ras85DV12.Scer\UAS-dlg1GD4689 MARCM third instar imaginal disc clones. Knockdown of either Pi3K92E or Akt1 in Ras85DV12.Scer\UAS-dlg1GD4689 cells, through expression of either Pi3K92EGD11228 or Akt1GD1361, causes a complete suppression of neoplastic growth and even causes a near-complete absence of mutant eye cells. Only small structures, which contain few cells, remain. MARCM Ras85DV12.Scer\UAS-dlg1GD4689 expressing clones are dramatically smaller in a Pi3K92E1C1 homozygous mutant than in a wild-type background, indicating that a loss of Pi3K92E signaling interferes with Ras85DV12.Scer\UAS-dlg1GD4689 tumour growth. Depletion of InR, through expression of InRGD104, slightly reduces tumour size in Ras85DV12.Scer\UAS-dlg1GD4689 MARCM clones. Depletion of S6k, through expression of S6kGD6646, slightly reduces tumour size in Ras85DV12.Scer\UAS-dlg1GD4689 MARCM clones. Depletion of Rheb, through expression of RhebNIG.1081R, strongly reduces tumour size in Ras85DV12.Scer\UAS-dlg1GD4689 MARCM clones. Depletion of Tor, through expression of TorNIG.5092R, strongly reduces tumour size in Ras85DV12.Scer\UAS-dlg1GD4689 MARCM clones. Depletion of Pten, through expression of PtenKK109278, increases tumour size in Ras85DV12.Scer\UAS-dlg1GD4689 MARCM clones. Overexpression of ykiScer\UAS.cHa in dlg1GD4689 MARCM clones generates larger tumorigenic overgrowths than in either single mutant clone. Expression of Ras85DV12.Scer\UAS and dlg1GD4689 in the posterior compartment of wing discs, under the control of Scer\GAL4en.PU results in an enlarged posterior wing disc compartment. These wing discs exhibit high levels of cell proliferation. Co-expression of Pi3K92EGD11228 together with Ras85DV12.Scer\UAS and dlg1GD4689, under the control of Scer\GAL4en.PU, strongly reduces the size of the Ras85DV12.Scer\UAS-dlg1GD4689 tumours in the posterior wing disc compartment. There is an almost complete absence of cellular proliferation in this compartment. Wing disc compartments expressing Ras85DV12.Scer\UAS, dlg1GD4689, Pi3K92EGD11228, together with CycEScer\UAS.cRa and stghs.PE2 show a rescue in compartment size and high levels of His3-positive cells.
Starting on day 8 after egg laying (AEL), Ras85DV12.Scer\UAS, Scer\GAL4Scer\FRT.Act5C tumours with a homozygous dorC107 background exhibit enhanced outgrowth when compared with Ras85DV12.Scer\UAS, Scer\GAL4Scer\FRT.Act5C controls. On day 14 AEL, these double mutant tumour cells clearly invade into the ventral nerve cord with a high frequency. On day 18 AEL, the dramatically overgrown double mutant tumours occupy about one-quarter volume of the larvae body and completely enveloped ventral nerve cord. Occasionally, tumour cells are also found in the gut and the trachea of the double mutant larvae, indicating secondary tumour formation in distal organs. All double mutant larvae die prior to pupation. Knockdown of car through expression of cardsRNA.Scer\UAS under the control of Scer\GAL4Scer\FRT.Act5C moderately enhances the overgrowth seen in Ras85DV12.Scer\UAS, Scer\GAL4Scer\FRT.Act5C tumour cells. Knockdown of Vps16A through expression of Vps16AdsRNA.Scer\UAS.WIZ under the control of Scer\GAL4Scer\FRT.Act5C moderately enhances the overgrowth seen in Ras85DV12.Scer\UAS, Scer\GAL4Scer\FRT.Act5C tumour cells and leads to tumour invasion towards the ventral nerve cord. Ras85DV12.Scer\UAS, Scer\GAL4Scer\FRT.Act5C tumours in a homozygous dorC107 background exhibit a massive accumulation of Hsap\LAMP1Scer\UAS.T:Avic\GFP-EGFP proteins in the leading edge of invading tumour cells indicating impaired lysosomal degradation. Expression of bskDN.Scer\UAS in Ras85DV12.Scer\UAS, Scer\GAL4Scer\FRT.Act5C /dorC107 clones suppresses the enhanced tumour overgrowth and metastasis.
scrib1 mutant eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU grow into large metastatic tumors. Wild type cells are almost completely absent from late Ras85DV12.Scer\UAS scrib1 tumors. Eye-antennal disc clones that express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU, and in which adjacent cells are mutant for scrib1, develop into large tumors capable of invading the ventral nerve cord. In the late stages of tumor development most cells in the tumor mass are Ras85DV12.Scer\UAS cells and scrib1 cells and wild type cells are almost completely absent. Eye-antennal disc clones that express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU, and in which adjacent cells are mutant for l(2)glunspecified, develop into large tumors capable of invading the ventral nerve cord. l(2)glunspecified mutant eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU grow into large metastatic tumors. Expression of upd1dsRNA.Scer\UAS.cUa partially suppresses the overgrowth and invasion of the nerve cord seen in l(2)glunspecified mutant eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. The phenotype is further suppressed when the clones are generated in a upd2Δ3-62 mutant background. upd2Δ3-62 does not suppress the overgrowth and invasion of the nerve cord seen in l(2)glunspecified mutant eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. Expression of domeΔCYT.Scer\UAS suppresses the overgrowth and invasion of the nerve cord seen in scrib1 mutant eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. Expression of domeΔCYT.Scer\UAS in eye-antennal disc clones that express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU, and in which adjacent cells are mutant for scrib1, suppresses the overgrowth and invasion of the nerve cord. Stat92E06346 partially suppresses the overgrowth and invasion of the nerve cord seen in scrib1 mutant eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. Stat92E06346 in eye-antennal disc clones that express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU, and in which adjacent cells are mutant for scrib1, partially suppresses the overgrowth and completely abrogates the invasion of the nerve cord. Eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS and upd1Scer\UAS.cUa under the control of Scer\GAL4Act.PU grow into large invasive tumors. Eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS and upd2Scer\UAS.cUa under the control of Scer\GAL4Act.PU grow into large invasive tumors. Eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS and upd3Scer\UAS.cUa under the control of Scer\GAL4Act.PU result in small non-invasive tumors. Eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS, upd1Scer\UAS.cUa and upd2Scer\UAS.cUa under the control of Scer\GAL4Act.PU grow into large invasive tumors that are larger than those seen when Ras85DV12.Scer\UAS is co-expressed with either upd1Scer\UAS.cUa or upd2Scer\UAS.cUa alone. Expression of domeΔCYT.Scer\UAS in eye-antennal disc clones that express Ras85DV12.Scer\UAS and upd1Scer\UAS.cUa under the control of Scer\GAL4Act.PU suppresses the overgrowth and invasion of the nerve cord. Expression of bskDN.Scer\UAS suppresses the tumors seen in scrib1 mutant eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU. Expression of bskDN.Scer\UAS does not suppress the tumors seen in eye-antennal disc clones that simultaneously express Ras85DV12.Scer\UAS and upd1Scer\UAS.cUa under the control of Scer\GAL4Act.PU. Expression of bskDN.Scer\UAS in eye-antennal disc clones that express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act.PU, and in which adjacent cells are mutant for scrib1, partially suppresses the tumor phenotype. Few scrib1 cells remain in the tissue at late stages.
Expression of Rab11S25N.Scer\UAS suppresses the enlarged lamina seen in third instar larval eye discs when Ras85DV12.Scer\UAS is expressed under the control of Scer\GAL4GMR.PF.
Hyperplasia in adult Malpighian tubule clones expressing Ras85DV12.Scer\UAS (using the MARCM system, under the control of Scer\GAL80αTub84B.PL and Scer\GAL4Scer\FRT.Act5C) is not suppressed by making them also mutant for aPKCk06403. Hyperplasia in adult Malpighian tubule clones expressing Ras85DV12.Scer\UAS (using the MARCM system, under the control of Scer\GAL80αTub84B.PL and Scer\GAL4Scer\FRT.Act5C) is suppressed by co-expression of Rho1N19.Scer\UAS. Hyperplasia in adult Malpighian tubule clones expressing Ras85DV12.Scer\UAS (using the MARCM system, under the control of Scer\GAL80αTub84B.PL and Scer\GAL4Scer\FRT.Act5C) is suppressed by co-expression of phlK497M.Scer\UAS. Hyperplasia in adult Malpighian tubule clones expressing Ras85DV12.Scer\UAS (using the MARCM system, under the control of Scer\GAL80αTub84B.PL and Scer\GAL4Scer\FRT.Act5C) is suppressed by co-expression of Dsor1VDRC.cUa. Hyperplasia in adult Malpighian tubule clones expressing Ras85DV12.Scer\UAS (using the MARCM system, under the control of Scer\GAL80αTub84B.PL and Scer\GAL4Scer\FRT.Act5C) is not suppressed by co-expression of Pi3K92EA2860C.Scer\UAS. Hyperplasia in adult Malpighian tubule clones expressing Ras85DV12.Scer\UAS (using the MARCM system, under the control of Scer\GAL80αTub84B.PL and Scer\GAL4Scer\FRT.Act5C) is not suppressed by making them also mutant for Stat92Ej6C8. Hyperplasia in adult Malpighian tubule clones expressing Ras85DV12.Scer\UAS (using the MARCM system, under the control of Scer\GAL80αTub84B.PL and Scer\GAL4Scer\FRT.Act5C) is suppressed by making them also mutant for Pka-C1B3. Hyperplasia in adult Malpighian tubule clones expressing Ras85DV12.Scer\UAS (using the MARCM system, under the control of Scer\GAL80αTub84B.PL and Scer\GAL4Scer\FRT.Act5C) is suppressed by making them also mutant for TorΔP.
Co-expression of RafK497M.UAS with Ras85DV12.UAS via Scer\GAL4NP7397 results in a phenotype similar to that of Ras85DV12.UAS overexpression alone. Mkp3UAS.cUa overexpression via Scer\GAL4NP7397 significantly suppresses the adult progenitor cell overproliferation phenotype induced by Ras85DV12.UAS in third instar larval midguts.
Expression of Ras85DV12.Scer\UAS within drkΔP24/+ adult mushroom bodies (under the control of either Scer\GAL4c747 or Scer\GAL4c739) results in severe learning deficits.
Ras85DV12.Scer\UAS Pten117 double mutant clones generated under the control of Scer\GAL4repo.PU are overgrown and invasive. Cells from mutant clones appear to invade the brain, typically following fibre tracts, and sometimes inducing the formation of trachea. Mutant cells often penetrate deep into the brain. Smaller clones are commonly seen that are composed of relatively differentiated, enlarged glia with diffusely invasive projections. Ras85DV12.Scer\UAS Pten117 double mutant optic lobe clones generated using eyFLP under the control of Scer\GAL4repo.PU contain more excess glial cells than in either mutant alone. The clones are composed of 10s of glial cells in third instar larvae, becoming large invasive tumors composed of hundreds to thousands of cells in adults.
The effects on wing shape and size in animals expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4MD-638 are partially rescued in a osa308/+ background, although these wings still show ectopic vein differentiation.
The wing notching that results from expressing Ras85DV12.Scer\UAS along the wing margin using Scer\GAL4vg.PU is partially suppressed by co-expression of picoIR.Scer\UAS.
Excessive photoreceptor differentiation can occur in mopT612 clones in the eye disc which are also expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4unspecified.
Co-expression of TorTED.Scer\UAS partially suppresses the salivary gland overgrowth phenotype that is seen in animals expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4fkh.PH and at 24 hours after puparium formation the salivary glands are predominantly degraded in the double mutant animals.
Clones in the eye antennal disc which are expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C and are also homozygous for scrib1 show invasive behaviour, invading the ventral nerve cord. These clones show degradation of the basement membrane. Co-expression of bskDN.Scer\UAS suppresses the invasive behaviour of clones in the eye antennal disc which are expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C and are also homozygous for scrib1. The invasive behaviour of clones in the eye antennal disc which are expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C and are also homozygous for scrib1 is partially suppressed in a Mmp1Q273 homozygous background. The invasive behaviour and degradation of the basement membrane of clones in the eye antennal disc which are expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C and are also homozygous for scrib1 is suppressed if the clones are also co-expressing both ReckScer\UAS.cSa and TimpScer\UAS.cSa simultaneously.
dlg114 mutant clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI exhibit invasive tumours. Expression of Ras85DV12.Scer\UAS in scrib1 mutant clones results in massive and metastatic tumours. After 6 days the cells exhibit moderate tumour growth and ventral nerve cord invasion phenotypes. Co-expression of bskDN.Scer\UAS and Ras85DV12.Scer\UAS in scrib1 mutant clones completely blocks the invasion of the ventral nerve cord as well as secondary tumour foci formation. The presence of an Akt11 mutant background considerably reduces the tumour load in scrib1 mutant clones expressing Ras85DV12.Scer\UAS (under the control of Scer\GAL4Act5C.PI) using the FLP/FRT system but does not impair metastatic behaviour. Overexpression of egrScer\UAS.cIa in Ras85DV12.Scer\UAS mutants (both under the control of Scer\GAL4Act5C.PI) using the FLP/FRT system in brain somatic clones results in accelerated tumour growth, despite nether expression of Ras85DV12.Scer\UAS or egrScer\UAS.cIa alone causing dramatic overgrowth. These tumorigenic regions do not invade into the ventral nerve cord. Expression of Traf4dsRNA.Scer\UAS (under the control of Scer\GAL4Act5C.PI) using the FLP/FRT system fails to suppress metastatic behaviour in l(2)gl4 mutants expressing Ras85DV12.Scer\UAS. Expression of Traf6dsRNA.Scer\UAS (under the control of Scer\GAL4Act5C.PI) using the FLP/FRT system strongly suppresses metastatic behaviour in l(2)gl4 mutants expressing Ras85DV12.Scer\UAS. Expression of Tak1dsRNA.Scer\UAS (under the control of Scer\GAL4Act5C.PI) using the FLP/FRT system strongly suppresses metastatic behaviour in l(2)gl4 mutants expressing Ras85DV12.Scer\UAS. Expression of hepdsRNA.Scer\UAS (under the control of Scer\GAL4Act5C.PI) using the FLP/FRT system strongly suppresses metastatic behaviour in l(2)gl4 mutants expressing Ras85DV12.Scer\UAS. Expression of wgndsRNA.Scer\UAS (under the control of Scer\GAL4Act5C.PI) using the FLP/FRT system partially suppresses metastatic behaviour in l(2)gl4 mutants expressing Ras85DV12.Scer\UAS. Expression of PvrdsRNA.Scer\UAS (under the control of Scer\GAL4Act5C.PI) using the FLP/FRT system does not suppress metastatic behaviour in l(2)gl4 mutants expressing Ras85DV12.Scer\UAS. shgk03401 mutant larval brain cells expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI exhibit a weak invasive phenotype and can invade the ventral nerve cord (this phenotype occurs with 20% penetrance). shgk03401 mutant larval brain cells expressing both Ras85DV12.Scer\UAS and egrScer\UAS.cIa under the control of Scer\GAL4Act5C.PI generate into invasive tumours that can invade the ventral nerve cord (this phenotype occurs with 88% penetrance). Expression of Ras85DV12.Scer\UAS, egrScer\UAS.cIa and bskDN.Scer\UAS, under the control of Scer\GAL4Act5C.PI in shgk03401 mutant larval cephalic complexes (brain, eye and antennal discs) induces tumor invasion of the ventral nerve cord. Overexpression of hepCA.Scer\UAS in shgk03401 mutant cephalic cells (i.e. cells in the larval brain and eye-antennal discs) expressing Ras85DV12.Scer\UAS results in neither enhanced tumour growth nor metastatic behaviour.
The roughened eye and increased rhabdomere phenotype seen when Ras85DV12.Scer\UAS is expressed under the control of Scer\GAL4hs.2sev is enhanced in a S6kIID1/+ or S6kIID1/Y background. These eye phenotypes are suppressed when either S6kIIScer\UAS.T:Hsap\MYC or S6kIIK231R.Scer\UAS.T:Hsap\MYC is coexpressed with Ras85DV12.Scer\UAS (all under the control of Scer\GAL4hs.2sev).
Coexpression of CycEScer\UAS.cLa and Ras85DV12.Scer\UAS, under the control of Scer\GAL4en-e16E, in the posterior compartment cells of embryos, leads to a reduction in compartment size, similar to that seen when only Ras85DV12.Scer\UAS is expressed, and cells are of a size similar to that seen when only CycEScer\UAS.cLa is expressed.
Mutant clones that express Ras85DV12.Scer\UAS, under the control of Scer\GAL4da.G32, in an ave108V background show the ave108V photoreceptor differentiation phenotype. The Scer\GAL4da.G32>Ras85DV12.Scer\UAS ectopic photoreceptor phenotype is therefore suppressed.
Clones of eye/antennal imaginal disc cells that express Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI and are also homozygous for scrib1 show an enhancement of the tumorigenic phenotype seen in clones that are mutant for either scrib1 or express Ras85DV12.Scer\UAS. The Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, scrib1 cells become invasive; at 6 days after egg laying, imaginal disc cells appear in the ventral nerve cord (VNC). At day 9, the invading cells obscure the eye/antennal discs, brain lobes, and the VNC so these anatomical structures are no longer distinguishable. The invading tumor cells are morphologically distinct and have a prominent actin cytoskeleton network typical of cells undergoing movement or shape change. Animals bearing Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, scrib1 clones progress normally through larval development but fail to pupate and die as third instar larvae and around day 13 AEL. Expression of bskDN.Scer\UAS clones blocks the cell invasive phenotype of Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, scrib1 eye/antennal imaginal disc clones. These clones still overgrow in the discs but never leave this structure. bskDN.Scer\UAS expression also rescues the larval lethality of Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, scrib1 clone-bearing animals, with rescued animals progressing through most of metamorphosis and dying as pharate adults. Expression of pucScer\UAS.cMa in Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, scrib1 eye/antennal imaginal disc clones prevents the invasion cell phenotype but does not rescue the overgrowth of the eye/antennal imaginal disc clones. pucScer\UAS.cMa expression rescues the larval lethality of Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, scrib1 clone-bearing animals and partially restores pupariation. Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, TimpScer\UAS.cPa, scrib1 eye/antennal disc clones produce a tumor mass larger than that seen in Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, scrib1 disc clones. However, the tumor cell invasiveness observed in Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, scrib1 clones is suppressed by expression of TimpScer\UAS.cPa. Expression of either Mmp1dsRNA.Scer\UAS or Mmp2dsRNA.Scer\UAS in Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, scrib1 eye/antennal imaginal disc clones significantly suppresses the cell invasiveness phenotype but does not suppress the formation of tumors within the disc. Coexpression of Mmp1dsRNA.Scer\UAS and Mmp2dsRNA.Scer\UAS further reduces the invasiveness of Scer\GAL4Act5C.PI>Ras85DV12.Scer\UAS, scrib1 cells. Coexpression of hepAct.Scer\UAS and Ras85DV12.Scer\UAS, under the control of Scer\GAL4Act5C.PI, in eye/antennal imaginal disc clones results in migration of some disc cells to ectopic regions of the brain, although cell invasion is not efficient in this genotype. Coexpression of hepScer\UAS.cBa and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI results in these discs results in the invasion of the disc cells into the ventral nerve cord and regions of the brain.
Ras85DV12.Scer\UAS; Scer\GAL4srp.Hemo suppresses the macrophage aggregation phenotype of stage 16 Pvr1 homozygous embryos. However, these embryos have a mild deficiency of hemocytes in the ventral-posterior region and in the posterior end of the elongated germ band.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4sim.P3.7 reduces the penetrance of the loss of midline glia phenotype seen in homozygous jing3 embryos, so that only 11.3% of double mutant embryos have a reduced number of midline glial cells.
Denticle belt fusions in the cuticles of rho7M43; ru1 double homozygous embryos are suppressed Ras85DV12.Scer\UAS; Scer\GAL4prd.RG1.
Scer\GAL4αTub84B.PL-driven expression of Ras85DV12.Scer\UAS in scrib1 clones leads to massive overproliferation of the scrib1 tissue, larval/pupal lethality. The addition of E2f191 or dapScer\UAS.cdNa suppresses the overproliferation phenotype seen in scrib1, Ras85DV12.Scer\UAS, Scer\GAL4αTub84B.PL clones.
Ras85DV12.Scer\UAS; Scer\GAL4twi.2PE substantially suppresses the visceral mesoderm phenotype seen in stage 12 jeb-c1 homozygous embryos.
Marked clones in the eye disc expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI and which are also homozygous for scribunspecified show metastatic behaviour; third instar larvae carrying these marked clones have large primary tumours in the head and also have small groups of cells floating in the hemolymph and other distant sites. The majority of ectopic tumour cells spread from the primary tumour onto the ventral nerve cord (VNC), eventually enveloping it and also spread into the first and second leg discs and tracheal vasculature at a lower frequency. The mutant cells can invade the inside of the VNC and the leading edge of the cells has a morphology common to actively migrating cells. The basement membrane has many points of discontinuity in eye discs containing clones which are homozygous for scribunspecified and which are expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI, and mutant cells spread from these areas. The basement membrane has many points of discontinuity in eye discs containing clones which are homozygous for scrib1 and which are expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI, and mutant cells spread from these areas. The metastatic behaviour seen in marked clones in the eye disc which are homozygous for scribunspecified and are also expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI is suppressed if they are co-expressing scribScer\UAS.cBa. Marked clones in the eye disc expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI and which are also homozygous for wtsunspecified result in immensely overgrown tumours in the eye-antennal discs, but marked cells are never seen on the ventral nerve cord or other locations in third instar larvae (indicating that the mutant tumour cells do not show metastatic behaviour). Marked clones in the eye disc expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI and which are also homozygous for scrib1 show metastatic behaviour. The metastatic behaviour seen in clones in the eye disc which are homozygous for scrib1 and are also expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI is suppressed if they are co-expressing shgScer\UAS.cOa but is not suppressed if they are co-expressing argos::shgi.Scer\UAS.T:Hsap\MYC. Marked clones in the eye disc expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI and which are also homozygous for shgunspecified do not show metastatic behaviour, although the cells cause disc eversion defects during metamorphosis. Marked clones in the eye disc expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI and which are also homozygous for one of baz4, sdtN5, Cdc421, l(2)gl4 or dlg114 show metastatic behaviour. The metastatic behaviour seen in clones in the eye disc which are homozygous for scrib1 and are also expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PI is not suppressed if they are co-expressing Hsap\CDKN1AScer\UAS.cHa; although the tumour size is decreased, mutant cells can still spread into the ventral nerve cord.
Clones expressing Ras85DV12.Scer\UAS under the control of Scer\GAL4αTub84B.PC usually do not stimulate the growth of vnL6/vnddd-4 wing discs. A few discs do show a modest increase in size, but this rescue appears to be due to a cell-autonomous rescue of proliferation within the Ras85DV12.Scer\UAS expressing clone.
Over 50% of flies expressing both Ras85DV12.Scer\UAS and spenΔC.Scer\UAS under the control of Scer\GAL4sli.PS are viable and appear to be normally patterned.
Expression of stgScer\UAS.cNa under the control of Scer\GAL4Act5C.PP in clones in the imaginal discs results in cells that have only slightly shorter doubling times than normal. Co-expression of both stgScer\UAS.cNa and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP results in shorter doubling times and cells of normal size. Large G1 and small G2 cell populations are seen (as is seen when stgScer\UAS.cNa is expressed under the control of Scer\GAL4Act5C.PP). The areas of clones co-expressing both stgScer\UAS.cNa and Ras85DV12.Scer\UAS under the control of Scer\GAL4Act5C.PP are increased compared to controls (as is seen when Ras85DV12.Scer\UAS is expressed under the control of Scer\GAL4Act5C.PP). The inhibitory effect on cell cycle rates of RbfScer\UAS.cDa expressed under the control of Scer\GAL4Act5C.PP in clones in the imaginal discs is not suppressed by co-expression of Ras85DV12.Scer\UAS. Co-expression of RbfScer\UAS.cDa and Ras85DV12.Scer\UAS has an additive effect on cell size.
When expression is driven in stumps09904b mutants by Scer\GAL4btl.PS or by Scer\GAL4hs.PB, many or all tracheal cells show erratic outgrowth of primary branches. These features are not normally seen in stumps09904b mutants. When expression is driven by Scer\GAL4twi.PG in stumps09904b embryos, some random mesodermal migrations are restored.
Co-expression of Ras85DV12.Scer\UAS and ovoD1.Scer\UAS driven by Scer\GAL4e22c or Scer\GAL469B suppresses the extra denticle phenotype seen by ubiquitous expression of Ras85DV12.Scer\UAS alone.
Photoreceptor differentiation and growth is strongly rescued in flies expressing wgScer\UAS.cAa under the control of Scer\GAL4ey.PH if they are also co-expressing Ras85DV12.Scer\UAS. The ectopic photoreceptor formation seen in clones expressing Ras85DV12.Scer\UAS in the eye disc under the control of Scer\GAL4Act5C.PP is not prevented by coexpression of armΔ.Scer\UAS.T:Ivir\HA1. eyg1/Df(3L)iro-2 hemizygous eye discs completely lack photoreceptors. Photoreceptor development in these discs is rescued by Ras85DV12.Scer\UAS expressed under the control of Scer\GAL4dpp.PH. The lack of photoreceptor phenotype of eya1 eye discs is not rescued by Ras85DV12.Scer\UAS expressed under the control of Scer\GAL4ey.PH.
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4btl.PS in btlLG19 or stumps09904b mutant embryos partially rescues tracheal morphogenesis.
Xenogenetic Interactions
Statement
Reference
Hsap\S100A4wt.UAS enhances the tumorigenesis of whole eye-antennal disc clones expressing Ras85DV12.UAS under the control of Scer\GAL4Act5C.PI, leading to extensive metastasis to the VNC and other organs, particularly to the gut and gonads. In contrast, Hsap\S100A4Δ2.UAS only results in a modest enhancement.
Co-expression of Hsap\FOXO1A::Hsap\PAX7Scer\UAS.cGa and Ras85DV12.Scer\UAS under the control of Scer\GAL4Mhc.PW results in disorganized larval musculature: the contours of many myofibres are vague and ill-defined. Collections of mononuclear cells are present (at the expense of syncitial myofiber tissue) at a significantly higher frequency than in larvae expressing Hsap\FOXO1A::Hsap\PAX7Scer\UAS.cGa alone.
Coexpression with BacA\p35Scer\UAS.cHa greatly reduces cell death but animals still die as young adults with deletion of structures, presumably as a consequence of non-autonomous cell death. Scer\GAL4dpp.blk1-mediated expression of one copy of Hsap\CDKN1AScer\UAS.cHa suppresses most, but not all, of Ras85DV12.Scer\UAS-induced cell proliferation and hyperplastic growth seen in wing discs. The lethal phase is delayed but not suppressed. Coexpression with two copies of Hsap\CDKN1AScer\UAS.cHa results in the complete suppression of lethality associated with Ras85D expression. Morphologically, the wing is restored to its normal shape and size, although Hsap\CDKN1AScer\UAS.cHa only partially suppresses the extra wing vein material along L3. Larger wing cell size is completely suppressed.
Complementation and Rescue Data
Comments
Expression of Ras85DV12.Scer\UAS under the control of Scer\GAL4αTub84B.PL in combination with Scer\GAL80ts.αTub84B in eye disc clones rescues the ommatidial development defects of Ras85DΔC40B homozygous cells. Clones show hyper-recruitment of photoreceptors.
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