Screen for mitochondrial DNA copy number maintenance genes reveals essential role for ATP synthase
Copyright policy )Screen for mitochondrial DNA copy number maintenance genes reveals essential role for ATP synthase
The machinery of mitochondrial DNA (mtDNA) maintenance is only partially characterized and is of wide interest due to its involvement in disease. To identify novel components of this machinery, plus other cellular pathways required for mtDNA viability, we implemented a genome-wide RNAi screen in Drosophila S2 cells, assaying for loss of fluorescence of mtDNA nucleoids stained with the DNA-intercalating agent PicoGreen. In addition to previously characterized components of the mtDNA replication and transcription machineries, positives included many proteins of the cytosolic proteasome and ribosome (but not the mitoribosome), three proteins involved in vesicle transport, some other factors involved in mitochondrial biogenesis or nuclear gene expression, > 30 mainly uncharacterized proteins and most subunits of ATP synthase (but no other OXPHOS complex). ATP synthase knockdown precipitated a burst of mitochondrial ROS production, followed by copy number depletion involving increased mitochondrial turnover, not dependent on the canonical autophagy machinery. Our findings will inform future studies of the apparatus and regulation of mtDNA maintenance, and the role of mitochondrial bioenergetics and signaling in modulating mtDNA copy number.
- Tampere University of Technology Finland
- University Federico II of Naples Italy
- Maastricht University Netherlands
- University of Campania "Luigi Vanvitelli" Italy
- Tampere University Finland
Medicine (General), mitochondrial biogenesis, complex V; DNA replication; mitochondrial biogenesis; mitochondrial DNA; mitophagy; nuclease; nucleoid; reactive oxygen species; Animals; Autophagy; Cell Line; DNA, Mitochondrial; Drosophila Proteins; Drosophila melanogaster; Gene Dosage; Gene Expression Regulation; Gene Library; Genome; Mitochondrial Proton-Translocating ATPases; RNA Interference; Reactive Oxygen Species; Genes, Essential, Gene Dosage, mitochondrial DNA, SACCHAROMYCES-CEREVISIAE, RNA-POLYMERASE, DNA replication; complex V; mitochondrial DNA; mitochondrial biogenesis; mitophagy; nuclease; nucleoid; reactive oxygen species, Drosophila Proteins, nuclease, Biology (General), reactive oxygen species, Genes, Essential, Genome, complex V, Articles, Mitochondrial Proton-Translocating ATPases, cell and molecular biology, BINDING-PROTEIN, Drosophila melanogaster, solu- ja molekyylibiologia - Biochemistry, OXYGEN SPECIES PRODUCTION, RNA Interference, 570, nucleoid, Biolääketieteet - Biomedicine, QH301-705.5, ENDOPLASMIC-RETICULUM, 610, DNA replication, DNA, Mitochondrial, Biokemia, solu- ja molekyylibiologia - Biochemistry, cell and molecular biology, Cell Line, R5-920, POLYMERASE-III SUBUNITS, Autophagy, Animals, DYNAMIN-RELATED GTPASE, Gene Library, TRANSCRIPTION FACTOR-A, Biokemia, CELLS LACKING, Biomedicine, mitophagy, Gene Expression Regulation, DOMAIN-CONTAINING 3A, Reactive Oxygen Species
Medicine (General), mitochondrial biogenesis, complex V; DNA replication; mitochondrial biogenesis; mitochondrial DNA; mitophagy; nuclease; nucleoid; reactive oxygen species; Animals; Autophagy; Cell Line; DNA, Mitochondrial; Drosophila Proteins; Drosophila melanogaster; Gene Dosage; Gene Expression Regulation; Gene Library; Genome; Mitochondrial Proton-Translocating ATPases; RNA Interference; Reactive Oxygen Species; Genes, Essential, Gene Dosage, mitochondrial DNA, SACCHAROMYCES-CEREVISIAE, RNA-POLYMERASE, DNA replication; complex V; mitochondrial DNA; mitochondrial biogenesis; mitophagy; nuclease; nucleoid; reactive oxygen species, Drosophila Proteins, nuclease, Biology (General), reactive oxygen species, Genes, Essential, Genome, complex V, Articles, Mitochondrial Proton-Translocating ATPases, cell and molecular biology, BINDING-PROTEIN, Drosophila melanogaster, solu- ja molekyylibiologia - Biochemistry, OXYGEN SPECIES PRODUCTION, RNA Interference, 570, nucleoid, Biolääketieteet - Biomedicine, QH301-705.5, ENDOPLASMIC-RETICULUM, 610, DNA replication, DNA, Mitochondrial, Biokemia, solu- ja molekyylibiologia - Biochemistry, cell and molecular biology, Cell Line, R5-920, POLYMERASE-III SUBUNITS, Autophagy, Animals, DYNAMIN-RELATED GTPASE, Gene Library, TRANSCRIPTION FACTOR-A, Biokemia, CELLS LACKING, Biomedicine, mitophagy, Gene Expression Regulation, DOMAIN-CONTAINING 3A, Reactive Oxygen Species
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