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General Information
Symbol
Dmel\shiK44A.UAS
Species
D. melanogaster
Name
FlyBase ID
FBal0101154
Feature type
allele
Associated gene
Associated Insertion(s)
Carried in Construct
Also Known As
UAS-shiK44A, UAS-shiDN, ShiDN, ShiK44A, UAS-shi.K44A, UAS-shiK44A, UAS-ShiD(K44A)
Key Links
Allele class
Nature of the Allele
Allele class
Mutations Mapped to the Genome
 
Type
Location
Additional Notes
References
Associated Sequence Data
DNA sequence
Protein sequence
 
 
Progenitor genotype
Carried in construct
Cytology
Nature of the lesion
Statement
Reference

A dominant negative form of shi (with a K44A mutation in the GTP-binding domain) 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 ( 0 )
Disease
Evidence
References
Modifiers Based on Experimental Evidence ( 0 )
Disease
Interaction
References
Comments on Models/Modifiers Based on Experimental Evidence ( 0 )
 
Disease-implicated variant(s)
 
Phenotypic Data
Phenotypic Class
Phenotype Manifest In
Detailed Description
Statement
Reference

Scer\GAL4He.PZ-driven expression of shiK44A.Scer\UAS does not induce increase in the number of circulating hemocytes in third instar larvae relative to controls.

Animals expressing shiK44A.Scer\UAS under the control of Scer\GAL4grh.D4 are lethal, show defects in larval dorsal trunk elongation (resulting in internalization of posterior spiracles into the body cavity) and the larvae are smaller than age-matched controls.

The adulthood-only expression of shiK44A.UAS under the control of Scer\GAL4esg-NP5130 (and Gal80[ts], for the temporal control of expression) leads to a significant increase in the number of intestinal stem cells and of mitotic figures in the adult midgut epithelium, as compared to controls.

Expression of shiK44A.Scer\UAS under the control of Scer\GAL4SPARC-MI00329-GAL4 results in elongation of the ventral nerve cord.

Third larval instar adipocytes expressing shiK44A.Scer\UAS under the control of Scer\GAL4SPARC-MI00329-GAL4 show a significant increase in plasma membrane depth compared to control cells.

Expression of shiK44A.Scer\UAS in the mushroom body, under the control of Scer\GAL4ey-OK107 results in mild structural defects in the mushroom body.

Expression of shiK44A.Scer\UAS under the control of either Scer\GAL4eg-Mz360, Scer\GAL4eve.RN2 or Scer\GAL4elav-C155 does not result in any defects in axon outgrowth in the embryo.

Early third instar larvae expressing shiK44A.Scer\UAS under the control of either Scer\GAL4tsh-Gal4-1, Scer\GAL4elav.PLu or Scer\GAL4Cha.7.4 at the restrictive temperature of 36[o]C are almost completely paralysed, with only occasional and highly aberrant muscle contractions.

Early third instar larvae expressing shiK44A.Scer\UAS under the control of "BL-Gal4" (a combination of Scer\GAL4elav.PLu, Scer\GAL80tsh-GAL80 and Scer\GAL80Cha.PK) at the restrictive temperature of 36[o]C continue to crawl actively, with forward waves of peristaltic contraction. The timing of contractions and the number of forward and backward waves are not significantly different from those of controls. There is no significant alteration in the number of pause turns compared to controls.

Early third instar larvae expressing shiK44A.Scer\UAS under the control of "BL+sens-Gal4" (a combination of Scer\GAL4elav.PLu and Scer\GAL80tsh-GAL80) at the restrictive temperature show defects in crawling. The larvae perform significantly fewer forward peristaltic waves and significantly more backward peristaltic waves than controls. The forward and backward waves are very slow. The larvae can perform pause turns, but their frequency is reduced compared to controls.

Expression of shiK44A.Scer\UAS under the control of Scer\GAL4twi.PG severely disturbs myoblast fusion in the embryo.

Expression of shiK44A.Scer\UAS under the control of either Scer\GAL4kirre-rP298 or Scer\GAL4sns.PK does not result in myoblast fusion defects in the embryo.

Primary macrophages isolated from animals expressing shiK44A.Scer\UAS under the control of Scer\GAL4Pxn.PS lack F-actin rich protrusions.

Expression of shiK44A.Scer\UAS under the control of Scer\GAL4Myo31DF-NP0001 (restricted to the larval stages using Scer\GAL80ts.αTub84B) results in defects in gut lipid mobilisation. At 3 hours neutral lipid droplets accumulate over a broader region of the second instar larval posterior midgut and their number within individual enterocytes is increased. By 6 hours most of the posterior midgut is filled with large lipid droplets. However, 24 hours after induction of shiK44A.Scer\UAS lipid droplets in the gut are strongly reduced.

Central nervous system neuroblasts of fed second instar larvae expressing shiK44A.Scer\UAS under the control of either Scer\GAL4Cg.PA or Scer\GAL4repo.PU show a reduction in EdU incorporation compared to controls.

Central nervous system neuroblasts of fed larvae expressing shiK44A.Scer\UAS under the control of Scer\GAL4n-syb.PS show no significant difference in EdU incorporation compared to controls.

Border cells expressing shiK44A.Scer\UAS under the control of Scer\GAL4slbo.2.6 show a reduced rate of uptake of the lipophilic dye FM4-64 compared to wild-type controls.

The luminal chitin extracellular matrix is disorganised and the gap region is lost in the tracheal tubes of embryos expressing shiK44A.Scer\UAS under the control of Scer\GAL4unspecified in the tracheal cells.

Expression of shiK44A.Scer\UAS, under the control of Scer\GAL4ptc-559.1 in first instar larvae results in the domain of type 2 cells in the epidermis being reduced, whereas the domain of type 3 cells is enlarged. This alters the cuticle bristle pattern.

Expression of shiK44A.Scer\UAS in the fat cells, under the control of Scer\GAL4Scer\FRT.Rnor\CD2.Act5C blocks internalisation of TR-avidin, resulting in the accumulation of large aggregates near the cell surface.

Expression of shiK44A.Scer\UAS under the control of Scer\GAL4Scer\FRT.Rnor\CD2.Act5C results in a 1.5-fold increase in fat body cell size, consistent with its growth effects in wing disc cells.

Disruption of endocytosis through expression of shiK44A.Scer\UAS (under the control of Scer\GAL4Scer\FRT.Rnor\CD2.Act5C) prevents the proper induction of autophagy after starvation.

Expression of shiK44A.Scer\UAS under the control of Scer\GAL4Scer\FRT.Rnor\CD2.Act5C results in increased cell size in imaginal discs.

In some embryos expressing shiK44A.Scer\UAS under the control of Scer\GAL4fkh.PH, the salivary gland does not complete its posterior migration.

Expression of shiK44A.Scer\UAS under the control of Scer\GAL4C96 or Scer\GAL4Bx-MS1096 causes apoptosis in the wing disc.

Few if any TUNEL-positive columnar cells are observed in shiK44A.Scer\UAS/Scer\GAL4bbg-C96 wing discs.

Few if any TUNEL-positive columnar cells are observed in shiK44A.Scer\UAS/Scer\GAL4C5 wing discs.

Expression of shiK44A.Scer\UAS under the control of Scer\GAL4bbg-C96 results in a loss of wing margin tissue.

Wings from shiK44A.Scer\UAS/Scer\GAL4C5 adults are small with altered morphology, such as bristle loss.

Expression of shiK44A.Scer\UAS under the control of Scer\GAL4ato.3.6 does not affect dorsal cluster neuron axon extension across the optic chiasm in adult flies.

Expression of shiK44A.Scer\UAS in wg-expressing cells, driven by Scer\GAL4wg.PM, in the embryos causes the wg protein to remain at wg-expressing cells. The formation of cluster I and cluster II slou founder cells in the mesoderm is not affected. Expression of shiK44A.Scer\UAS throughout the mesoderm, driven by Scer\GAL4twi.PB, blocks endocytosis and causes cluster II slou founder cells to be expanded in 53% of hemisegments.

Expression of shiK44A.Scer\UAS in border cells, driven by Scer\GAL4slbo.2.6 inhibits the migration of border cell clusters to the oocyte. However, it does not block the formation of the egg chamber's apical cap.

Expression of shiK44A.Scer\UAS, under the control of Scer\GAL4slbo.2.6, induces a border cell migration defect in 95% of egg chambers.

About 60% of stage 16 shiK44A.Scer\UAS; Scer\GAL4sim.PS embryos have missing or reduced ventral nerve cord commissures. When Scer\GAL4rho.PL is used as a driver the penetrance of this phenotype rises to 100%.

Expression of shiK44A.Scer\UAS under the control of Scer\GAL4Tab2-201Y does not result in mushroom body defects. Expression of shiK44A.Scer\UAS under the control of Scer\GAL4OK107 results in gross defects in the whole brain.

When expressed using a uniform Scer\GAL4 driver, P{UAS-shi.K44A}3-10 provides a very high level of endocytotic disruption, comparable to the original shi1 mutation. It phenocopies the cell-lethal effect of shi1. When expressed together using a uniform Scer\GAL4 driver, P{UAS-shi.K44A}4-1 and P{UAS-shi.K44A}3-7 provide a moderate level of endocytotic disruption. Separately they each provide a low level of disruption.

High level or ubiquitous expression of shiK44A.Scer\UAS (using an Scer\GAL4 driver) abolishes cuticle deposition. When shiK44A.Scer\UAS is expressed at moderate levels under the control of Scer\GAL4prd.RG1, vesicles are occasionally seen in the Scer\GAL4prd.RG1 expression domain, indicating that moderate expression of shiK44A.Scer\UAS reduces, but does not completely abolish, endocytosis. The embryos have a smaller expanse of naked cuticle in odd-numbered segments than is normal. Ectopic denticles appear at the posterior edge of the odd-numbered denticle belts. Denticle morphology in the even-numbered denticle belts is disrupted; the denticles are still organised in a six-row belt, but the anterior rows do not show the distinct morphologies characteristic of wild type. When shiK44A.Scer\UAS is expressed at high levels under the control of Scer\GAL4prd.RG1 defective cuticle deposition is seen, with the resulting holes in the cuticle corresponding roughly to the Scer\GAL4prd.RG1 expression domain. Expression of shiK44A.Scer\UAS under the control of Scer\GAL4wg.PM does not affect the cuticular pattern. Expression of shiK44A.Scer\UAS under the control of Scer\GAL4en-e16E perturbs denticle type specification in the first three rows of denticles in each segment. Expression of shiK44A.Scer\UAS under the control of Scer\GAL4en-e16E does not disrupt embryonic head development. Expression of high levels of shiK44A.Scer\UAS under the control of Scer\GAL4en-e16E results in severe segmental defects in cuticle deposition. Expression of shiK44A.Scer\UAS under the control of Scer\GAL4e22c results in cell death and a failure to secrete cuticle.

External Data
Interactions
Show genetic interaction network for Enhancers & Suppressors
Phenotypic Class
Phenotype Manifest In
Suppressed by
Enhancer of
Suppressor of
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Additional Comments
Genetic Interactions
Statement
Reference

Flies expressing X11LdsRNA.Scer\UAS.cGa and shiK44A.Scer\UAS in the mushroom body, under the control of Scer\GAL4ey-OK107, are embryonic lethal, indicating a robust genetic interaction between X11L and shi. Raising these flies at a lower temperature (so reducing expression of these transgenes) results in viable flies, displaying aberrant anatomical development of the medial and dorsal lobes.

Early third instar larvae expressing shiK44A.Scer\UAS under the control of "BL-Gal4" (a combination of Scer\GAL4elav.PLu, Scer\GAL80tsh-GAL80 and Scer\GAL80Cha.PK) at the restrictive temperature and also carrying WGMR.PG show a normal avoidance response to a spot of intense light.

Expression of shiK44A.Scer\UAS (under the control of Scer\GAL4ey.PH) strongly enhances the Torey.hs.T:Zzzz\FLAG phenotype.

Co-expression of shiK44A.Scer\UAS partially restores salivary gland integrity in embryos expressing Rac1V12.Scer\UAS under the control of Scer\GAL4fkh.PH, but the glands are narrower and have fewer cells than wild type.

Expression of shiK44A.Scer\UAS, under the control of Scer\GAL4twi.PB, in a wgl-17 /+ background suppresses the expansion of cluster II slou founder cells that occurs when the background is wild type.

Fusion of ventral nerve cord commissures seen in sli550 homozygous embryos is largely suppressed by shiK44A.Scer\UAS; Scer\GAL4rho.PL.

The phenotype produced when shiK44A.Scer\UAS is expressed at moderate levels under the control of Scer\GAL4prd.RG1 can be rescued by co-expression of wgScer\UAS.cHa; the naked cuticle region in odd-numbered segments is restored to its normal expanse and the even-numbered denticle belts show a more normal degree of diversity. The denticle phenotype caused by expression of shiK44A.Scer\UAS under the control of Scer\GAL4en-e16E can be rescued by co-expression of wgScer\UAS.cHa. Homozygous nkd2 embryos have excess naked cuticle replacing denticle belts in most segments and also have severe head defects. The formation of excess naked cuticle is suppressed by the expression of shiK44A.Scer\UAS under the control of Scer\GAL4en-e16E and the head defect is partially rescued in these embryos.

Xenogenetic Interactions
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Complementation and Rescue Data
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Mutant
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Stocks (4)
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External Crossreferences and Linkouts ( 0 )
Synonyms and Secondary IDs (10)
References (63)