title: Modulation of mitochondrial transcription in response to mtDNA depletion and repletion in HeLa cells.
abstract: The steady-state amounts of mitochondrial transcripts and transcription proteins were analyzed during mtDNA depletion and subsequent repletion to gain insight into the regulation of human mitochondrial gene expression. As documented previously, HeLa cells depleted of mtDNA via treatment with ethidium bromide (EB) were found to contain reduced steady-state levels of the mitochondrial transcription factor h-mtTFA. When partially mtDNA-depleted cells were cultured in the absence of EB, h-mtTFA recovered to normal levels at a significantly slower rate than mtDNA. Human mtRNA polymerase exhibited a similar depletion-repletion profile, suggesting that the mitochondrial transcription machinery is coordinately regulated in response to changes in mtDNA copy number. Newly synthesized mitochondrial transcripts were detected early in the recovery phase, despite the fact that mtDNA, h-mtTFA and h-mtRNA polymerase were simultaneously depleted. Although delayed relative to mtDNA, the amounts of h-mtTFA and h-mtRNA polymerase sharply increased during the later stages of the recovery phase, which was accompanied by accelerated rates of transcription and mtDNA replication. Altogether, these data indicate that when mtDNA copy number is low, it is beneficial to prevent accumulation of mitochondrial transcription proteins. In addition, h-mtTFA and h-mtRNA polymerase are either normally present in excess of the amount required for transcription or their activity is up-regulated to ensure continued expression and transcription-dependent replication of the mitochondrial genome during mtDNA-depleted states.
authors.affiliation: Department of Biochemistry, Rollins Research Center, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Bonnie L
authors.initials: BL
authors.lastname: Seidel-Rogol
authors.forename: Gerald S
authors.initials: GS
authors.lastname: Shadel
mesh_terms: D004261:DNA Replication; D004272:DNA, Mitochondrial / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004996:Ethidium / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008928:Mitochondria / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000096:biosynthesis / Q000235:genetics*; D009687:Nuclear Proteins; D012313:RNA / Q000096:biosynthesis*; D000077278:RNA, Mitochondrial; D014157:Transcription Factors / Q000096:biosynthesis; D014158:Transcription, Genetic; D015854:Up-Regulation; D020741:rho GTP-Binding Proteins / Q000235:genetics
chemical_list: D004272:DNA, Mitochondrial; D004268:DNA-Binding Proteins; D024101:Mitochondrial Proteins; D009687:Nuclear Proteins; D000077278:RNA, Mitochondrial; C499631:TFAM protein, human; D014157:Transcription Factors; C422562:mitochondrial transcription factor A; D012313:RNA; D012321:DNA-Directed RNA Polymerases; D020741:rho GTP-Binding Proteins; D004996:Ethidium