G12V | Ras85D-PA
Analogous to oncogenic mutations in one of three human RAS genes; mutation carried on in vitro construct; site of nucleotide substitution in fly gene inferred by FlyBase curator based on reported amino acid change.
This somatic variant has been found associated with cancer in each of the three paralogous genes in human (KRAS, HRAS, NRAS).
Co-expressing PtendsRNA.Exel.UAS and Ras85DV12.cUa.UAS under the control of Scer\GAL4repo.PU induces larval central brain gliomas; brain lobes are enlarged; there is an increase in mitosis; there is an increase in the number of neuroblasts; neuroblasts and ganglion mother cells seem to undergo a lateral shift at the edge of the central brain abutting the optic lobe.
Whole-eye disc clones both l(2)gl4 homozygous and expressing Ras85DV12.cUa.UAS under the control of Scer\GAL4Act5C.PP form tumors that invade into the VNC; the VNC invasion phenotype is nearly suppressed by the co-expression of pucUAS.cUa, partially suppressed by the co-expression of licJF01433 and partially suppressed by licG0252 heterozygosity; however, the size of the primary tumor is not affected by these manipulations.
Quadruple or quintuple co-expression for one week during adulthood of Ras85DV12.cUa.UAS, p53VDRC.cUa, PtenGD13500, ApcVDRC.cUa and/or MedVDRC.cUa under the combined control of Scer\GAL4byn-Gal4, Dicer-2 (for efficient RNAi) and Gal80[ts] (for the temporal control of expression) leads to multilayered epithelia that form bulges at discrete points along the hindgut (only in animals that contained Ras85DV12.cUa.UAS and ApcVDRC.cUa together) and to an expanded pylorus (observed in the quintuple and all quadruple conditions that carried Ras85DV12.cUa.UAS); the quadruple co-expression conditions Ras85DV12.cUa.UAS+p53VDRC.cUa+PtenGD13500+MedVDRC.cUa and p53VDRC.cUa+PtenGD13500+ApcVDRC.cUa+MedVDRC.cUa do not lead to multilayering.
All quadruple and quintuple co-expression conditions - with the exception of the quadruple p53VDRC.cUa+PtenGD13500+MedVDRC.cUa+ApcVDRC.cUa - result in numerous hindgut cells losing their characteristic epithelial shape to assume a more mesenchymal appearance (processes extending towards the basement membrane and surrounding muscle layer), migrate locally and disseminate to distant sites in the organism (e.g. head, legs, fat body, ovaries, below the epidermis of the abdomen and within the abdominal cavity). Similar hindgut cell dissemination is observed upon the triple expressions of Ras85DV12.cUa.UAS+p53VDRC.cUa+PtenGD13500, Ras85DV12.cUa.UAS+ApcVDRC.cUa+PtenGD13500 or Ras85DV12.cUa.UAS+p53VDRC.cUa+ApcVDRC.cUa, the double expressions of Ras85DV12.cUa.UAS+p53VDRC.cUa, Ras85DV12.cUa.UAS+PtenGD13500 or Ras85DV12.cUa.UAS+ApcVDRC.cUa, all of which are more severe than upon the single expression of Ras85DV12.cUa.UAS alone.
Somatic clones co-expressing wupAf06492 and Ras85DV12.cUa.Scer\UAS under the control of Scer\GAL4Act.PU in wandering third instar larval wing discs present a significant increase in the relative clonal area as compared to clones only expressing Ras85DV12.cUa.Scer\UAS, but not to clones only expressing wupAf06492, and lead to overgrowths protruding from the wing disc in almost half of cases.
The induction of somatic clones co-expressing l(2)glGD4047 and Ras85DV12.cUa.Scer\UAS under the control of Scer\GAL4Act.PU during second instar larval stage results in the accumulation of clonal cells in the wandering third instar larval fat body, which is enhanced by the additional co-expression of wupAJF02172; the induction of these clones also results in lethality before adulthood, which is partially rescued by the additional co-expression of wupAJF02172, although the surviving adults display a shorter life-span as compared to wild-type controls.
In wandering third instar larvae, somatic clones co-expressing l(2)glGD4047 and Ras85DV12.cUa.Scer\UAS under the control of Scer\GAL4Act.PU in both eye discs and wing discs exhibit a significant increase in the relative clonal area and hyperplasia as compared to control clones; wing disc clone cells localize both basally and apically in the disc. All of these phenotypes are suppressed by the additional co-expression of wupAJF02172, even leading to a significant decrease in the wing disc clonal area as compared to control clones.
Somatic clones co-expressing l(2)glGD4047, Ras85DV12.cUa.Scer\UAS and wupAf06492 under the control of Scer\GAL4Act.PU in wandering third instar larvae wing discs exhibit a significant increase in relative clonal area as compared to clones only co-expressing l(2)glGD4047 and Ras85DV12.cUa.Scer\UAS or only expressing wupAf06492; animals bearing clones co-expressing l(2)glGD4047, Ras85DV12.cUa.Scer\UAS and wupAf06492 do not reach adulthood.
Co-expression of Ras85DV12.cUa.Scer\UAS increases the extent of migration into the posterior compartment of anterior cells expressing Vha44Scer\UAS.T:Ivir\HA1 under the control of Scer\GAL4ptc-559.1 in the wing disc.
The triple co-expression of Ras85DV12.cUa.Scer\UAS, Hsap\UBE3AScer\UAS.cRa and either HPV18\E6Scer\UAS.T:Hsap\MYC or HPV18\E6V158A.Scer\UAS.T:Hsap\MYC, under the control of Scer\GAL4GMR.PU, leads to the migration of a considerable proportion of expressing eye tissue cells into distant pupal tissues (e.g. abdomen), as compared to controls. The triple co-expression of Ras85DV12.cUa.Scer\UAS, Hsap\UBE3AScer\UAS.cRa and HPV18\E6V158A.Scer\UAS.T:Hsap\MYC under the control of Scer\GAL4GMR.PU does not suppress the pupal lethality induced by the single expression of Ras85DV12.cUa.Scer\UAS under the control of Scer\GAL4GMR.PU.