UASp regulatory sequences drive expression of XhoI that is tagged at the N-terminal end with the Tag:Mito(kdn) mitochondrial targeting sequence. The protein is also tagged at the C-terminal end with Tag:MYC.
visible, with Scer\GAL4ey.PB
eye, with Scer\GAL4ey.PB
Expression of Xvas\XhoIUASp.Tag:Mito(kdn),Tag:MYC under the control of Scer\GAL4tim.PE (constitutively or in combination with tub-Gal80ts to restrict expression to the adult stage) leads to a significant reduction in the amplitude of locomotor activity rhythm (circadian rhythm) while its expression under the control of Scer\GAL4Pdf.PU does not lead to any changes in the amplitude of locomotor activity rhythm under constant darkness when compared to control adults. Also, expression of Xvas\XhoIUASp.Tag:Mito(kdn),Tag:MYC under the control of Scer\GAL4tim.PE leads to a significant decline of mitochondrial DNA in the adult head when compared to controls.
Expression of Xvas\XhoIScer\UAS.P\T.T:Mito-kdn,T:Hsap\MYC is embryonic/first instar larval lethal under the Scer\GAL4da.PU driver and pupal lethal under the Scer\GAL4VGlut-OK371 driver and the number of active zones in the neuromuscular junctions (NMJs) in third instar larvae is significantly decreased (while the number of boutons is not changed). The number and volume of presynaptic mitochondria at the NMJ is also strongly reduced relative to controls, however the ultrastructure of these mitochondria appear normal. Xvas\XhoIScer\UAS.P\T.T:Mito-kdn,T:Hsap\MYC expression affects mitochondria in other parts of the motor neuron as well: the cell body mitochondria are less reticular with more punctate morphology and both the number and volume of mitochondria in the distal axon is reduced, in the proximal axon the number of mitochondria is decreased but their volume is comparable to controls.
95% of animals expressing Xvas\XhoIScer\UAS.P\T.T:Mito-kdn,T:Hsap\MYC under the control of Scer\GAL4αTub84B.PL fail to hatch, and the remaining 5% die as first instar larvae.
Expression of Xvas\XhoIScer\UAS.P\T.T:Mito-kdn,T:Hsap\MYC under the control of Scer\GAL4ey.PB results in ablated or greatly reduced eyes.
Scer\GAL4ey.PB, Xvas\XhoIUASp.Tag:Mito(Cs1),Tag:MYC has visible phenotype, suppressible by mt:CoIA302T
Scer\GAL4ey.PB, Xvas\XhoIUASp.Tag:Mito(Cs1),Tag:MYC has visible phenotype, suppressible by mt:CoIR301Q
Scer\GAL4ey.PB, Xvas\XhoIUASp.Tag:Mito(Cs1),Tag:MYC has visible phenotype, suppressible by mt:CoIR301S
Scer\GAL4ey.PB, Xvas\XhoIUASp.Tag:Mito(Cs1),Tag:MYC has eye phenotype, suppressible by mt:CoIA302T
Scer\GAL4ey.PB, Xvas\XhoIUASp.Tag:Mito(Cs1),Tag:MYC has eye phenotype, suppressible by mt:CoIR301Q
Scer\GAL4ey.PB, Xvas\XhoIUASp.Tag:Mito(Cs1),Tag:MYC has eye phenotype, suppressible by mt:CoIR301S
The eye ablation phenotype caused by expression of Xvas\XhoIScer\UAS.P\T.T:Mito-kdn,T:Hsap\MYC under the control of Scer\GAL4ey.PB is completely suppressed if the animals also carry any one of mt:CoIA302T, mt:CoIR301Q or mt:CoIR301S.