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Saccharomyces cerevisiae Mhr1 can bind Xho I-induced mitochondrial DNA double-strand breaks in vivo.

Mitochondrion(2018)

Cited 5|Views18
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Abstract
Mitochondrial DNA (mtDNA) double-strand break (DSB) repair is essential for maintaining mtDNA integrity, but little is known about the proteins involved in mtDNA DSB repair. Here, we utilize Saccharomyces cerevisiae as a eukaryotic model to identify proteins involved in mtDNA DSB repair. We show that Mhr1, a protein known to possess homologous DNA pairing activity in vitro, binds to mtDNA DSBs in vivo, indicating its involvement in mtDNA DSB repair. Our data also indicate that Yku80, a protein previously implicated in mtDNA DSB repair, does not compete with Mhr1 for binding to mtDNA DSBs. In fact, C-terminally tagged Yku80 could not be detected in yeast mitochondrial extracts. Therefore, we conclude that Mhr1, but not Yku80, is a potential mtDNA DSB repair factor in yeast.
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Key words
mtDNA,DSB,oxidative phosphorylation,nuclear DNA,reactive oxygen species,homologous recombination,classical non-homologous end joining,yeast Ku80,Mhr1,yeast Ku,mitochondrial-targeted restriction endonuclease Xho I,red fluorescent protein,complete supplement mixture lacking uracil,complete supplement mixture lacking tryptophan,galactose,total cell extracts,Raff
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