Open AccessScreen for mitochondrial DNA copy number maintenance genes reveals essential role for ATP synthase
Author(s) -
Fukuoh Atsushi,
Cannino Giuseppe,
Gerards Mike,
Buckley Suzanne,
Kazancioglu Selena,
Scialo Filippo,
Lihavainen Eero,
Ribeiro Andre,
Dufour Eric,
Jacobs Howard T
Publication year - 2014
Publication title -
molecular systems biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 8.523
H-Index - 148
ISSN - 1744-4292
DOI - 10.15252/msb.20145117
Subject(s) - university hospital , library science , gene technology , biology , computer science , medicine , family medicine , microbiology and biotechnology
The machinery of mitochondrial DNA (mt DNA ) 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 mt DNA viability, we implemented a genome‐wide RNA i screen in Drosophila S2 cells, assaying for loss of fluorescence of mt DNA nucleoids stained with the DNA ‐intercalating agent PicoGreen. In addition to previously characterized components of the mt DNA 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 mt DNA maintenance, and the role of mitochondrial bioenergetics and signaling in modulating mt DNA copy number.