Screen for mitochondrial DNA copy number maintenance genes reveals essential role for ATP synthase
Fukuoh, Atsushi; Cannino, Guiseppe; Gerards, Mike; Buckley, Suzanne; Kazancioglu, Selena; Scialo, Filippo; Lihavainen, Eero; Ribeiro, Andre; Dufour, Eric; Jacobs, Howard T (2014)
Fukuoh, Atsushi
Cannino, Guiseppe
Gerards, Mike
Buckley, Suzanne
Kazancioglu, Selena
Scialo, Filippo
Lihavainen, Eero
Ribeiro, Andre
Dufour, Eric
Jacobs, Howard T
2014
Molecular Systems Biology 10 6
734
BioMediTech - BioMediTech
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi:uta-201610122432
https://urn.fi/URN:NBN:fi:uta-201610122432
Tiivistelmä
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.
Kokoelmat
- Artikkelit [6140]