UAS regulatory sequences drive expression of Hsap\MARK1 (codon-optimised for Drosophila, 781aa), tagged at the N-terminal with Tag:MYC and at the C-terminal with Tag:HA.
Flies expressing Hsap\MARK1Scer\UAS.T:Hsap\MYC,T:Ivir\HA1 under the control of Scer\GAL4VGlut-OK371 do not show any significant climbing ability defects relative to age-matched controls.
Hsap\MARK1UAS.Tag:MYC,Tag:HA, Hsap\TTBK1UAS.Tag:MYC,Tag:HA, Scer\GAL4nSyb.PS has short lived phenotype, enhanceable by Hsap\MAPTUAS.2N4R.cFa, Scer\GAL4nSyb.PS
Hsap\MAPTUAS.2N4R.cFa, Hsap\MARK1UAS.Tag:MYC,Tag:HA, Scer\GAL4nSyb.PS has short lived phenotype, enhanceable by Hsap\TTBK1UAS.Tag:MYC,Tag:HA, Scer\GAL4nSyb.PS
Hsap\MARK1UAS.Tag:MYC,Tag:HA, Hsap\TTBK1UAS.Tag:MYC,Tag:HA, Hsap\TTBK2UAS.Tag:MYC,Tag:HA, Scer\GAL4nSyb.PS has short lived phenotype, non-enhanceable by Hsap\MAPTUAS.2N4R.cFa, Scer\GAL4nSyb.PS
Hsap\MARK1UAS.Tag:MYC,Tag:HA, Scer\GAL4nSyb.PS is an enhancer of short lived phenotype of Hsap\TTBK1UAS.Tag:MYC,Tag:HA, Scer\GAL4nSyb.PS
Hsap\MAPTUAS.2N4R.cFa, Hsap\MARK1UAS.Tag:MYC,Tag:HA, Scer\GAL4nSyb.PS has short lived phenotype
Adult flies co-expressing both Hsap\MARK1Scer\UAS.T:Hsap\MYC,T:Ivir\HA1 and Hsap\MAPTScer\UAS.2N4R.cFa under the control of Scer\GAL4nSyb.PS combined with tub-Gal80[ts] to limit the expression to adulthood show significantly reduced lifespan relative to controls. Similarly, flies simultaneously expressing both these transgenes under the Scer\GAL4VGlut-OK371 driver display age-progressive locomotion deficit (climbing ability defects) that start to be apparent at day 7 post-eclosion.
Hsap\TTBK1Scer\UAS.T:Hsap\MYC,T:Ivir\HA1, Hsap\TTBK2Scer\UAS.T:Hsap\MYC,T:Ivir\HA1, Hsap\MAPTScer\UAS.2N4R.cFa and Hsap\MARK1Scer\UAS.T:Hsap\MYC,T:Ivir\HA1 expressed under the control of Scer\GAL4nSyb.PS combined with tub-Gal80[ts] to limit the expression to adulthood show combinatorial toxicity and lead to short adult lifespan. The short lifespan of adults expressing any two of the above mentioned transgenes is reduced even further when a third one is added (except for simultaneous expression of Hsap\TTBK1Scer\UAS.T:Hsap\MYC,T:Ivir\HA1 and Hsap\TTBK2Scer\UAS.T:Hsap\MYC,T:Ivir\HA1 where the combination with Hsap\MAPTScer\UAS.2N4R.cFa partially alleviates the reduced survival). When Hsap\TTBK1Scer\UAS.T:Hsap\MYC,T:Ivir\HA1, Hsap\TTBK2Scer\UAS.T:Hsap\MYC,T:Ivir\HA1, and Hsap\MARK1Scer\UAS.T:Hsap\MYC,T:Ivir\HA1 are co-expressed, the addition of Hsap\MAPTScer\UAS.2N4R.cFa cannot further shorten the adult lifespan.