Amino acid replacement: T231D.
Amino acid replacement: K250A.
The endogenous Pi3K92E promoter drives expression of a genomic fragment of Pi3K92E containing four site-directed mutations which alter four residues in the Ras-binding domain (RBD) of the Pi3K92E protein, thereby abolishing its ability to bind the Ras85D protein. The mutant protein is tagged with Tag:MYC.
Amino acid replacement: R253A.
Amino acid replacement: K257A.
Pi3K92Et.RBD.Tag:MYC partially rescues Pi3K92EA
The P{Myc-Dp110RBD} transgene rescues Pi3K92EA lethality, as assayed in "Df(3R)Pi3K92EA, P{HBS}/Df(3R)Pi3K92EA" mutants. The rescued flies are fertile and produce viable offspring. P{Myc-Dp110RBD} also rescues Pi3K92EA lethality when Pi3K21B dosage is halved, as assayed in "Df(2L)Pi3K21B-A/+; Df(3R)Pi3K92EA, P{HBS}, P{U2af38.gR10}/Df(3R)Pi3K92EA" mutants.
Pi3K92EA homozygous null flies (genotype Df(3R)Pi3K92EA, P{HBS}) rescued with 2 copies of the P{Myc-Dp110RBD} transgene have wings approximately 86% of the area of control flies (Pi3K92EA homozygous null flies rescued with 2 copies of the P{Myc-Dp110} transgene), while their body mass is reduced to approximately 69% of controls.
Pi3K92EA homozygous null flies (genotype Df(3R)Pi3K92EA, P{HBS}) rescued with 2 copies of the P{Myc-Dp110RBD} transgene lay fewer eggs than controls (Pi3K92EA homozygous null flies rescued with 2 copies of the P{Myc-Dp110} transgene) at ages. Consistent with this, Pi3K92EA homozygous null flies rescued with 2 copies of P{Myc-Dp110RBD} have smaller ovaries with fewer ovarioles than controls.