UASt regulates expression of a "mtDNA mutator" construct based on a mitochondrially-targeted cytidine deaminase, APOBEC1. The Rnor\Apobec1 cDNA is placed downstream of the mitochondrial targeting sequence of human ATP synthase subunit F1β (Tag:Mito(hATP5F1B)), a Tag:HA epitope and a flexible linker sequence 5'. A nuclear exporting sequence (NES) is inserted 3' to Rnor\Apobec1.
Expression of Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA under the control of Scer\GAL4da.G32 leads to an impaired climbing ability and a shortened lifespan in young flies when compared to controls.
The expression of Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA under the control of Scer\GAL4da.G32 does not induce any gross development or viability defect, but drastically shortens lifespan and climbing ability. Adults show higher levels of mitochondrial DNA (mtDNA) mutations, although indels and larger deletions are rare, there is no/minimal impact on mtDNA levels or mtDNA integrity; almost all mutations are C:G > T:A transitions; most of the cytidine deamination events occur in the minor strand; the frequency of mutations does not change with age; there is a significantly higher NS:S mutation ratio and mutations disproportionally target the 1st and 2nd codons in protein-coding genes, as compared to controls.
Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA, Scer\GAL4da.G32 has short lived phenotype, enhanceable by park25
Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA, Scer\GAL4da.G32 has short lived phenotype, enhanceable by StingΔRG5
Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA, Scer\GAL4da.G32 has abnormal locomotor behavior | adult stage phenotype, enhanceable by park25/StingΔRG5
Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA, Scer\GAL4da.G32, park25 has short lived phenotype, enhanceable by StingΔRG5
Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA, Scer\GAL4da.G32 has abnormal locomotor behavior | adult stage phenotype, non-enhanceable by park25
Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA, Scer\GAL4da.G32 has abnormal locomotor behavior | adult stage phenotype, non-enhanceable by StingΔRG5
Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA, Scer\GAL4da.G32 has abnormal locomotor behavior | adult stage phenotype, non-suppressible by park25
Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA, Scer\GAL4da.G32 has abnormal locomotor behavior | adult stage phenotype, non-suppressible by StingΔRG5
In young flies, StingΔRG5 and park25 mutations together enhance the impaired climbing ability phenotype observed as a result of expression of Rnor\Apobec1Tag:Mito(hATP5F1B),Tag:HA under the control of Scer\GAL4da.G32, while StingΔRG5 or park25 mutation alone does not modify that phenotype.