ArticleThe Journal of biological chemistry2010
Identification of multiple rate-limiting steps during the human mitochondrial transcription cycle in vitro.
Article in The Journal of biological chemistry, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Deciphering HIV-1 Transcription Initiation and Elongation from Single-Molecule Imaging Data.Research (Washington, D.C.) · 2025Article
- 35 Years of TFAM Research: Old Protein, New Puzzles.Biology · 2023Review
- Mechanisms of mammalian mitochondrial transcription.Protein science : a publication of the Protein Society · 2019Review
- Roles of the mitochondrial genetics in cancer metastasis: not to be ignored any longer.Cancer metastasis reviews · 2018Review
- SimpleAntimicrobial agents and chemotherapy · 2018Article
- Article
- Topological requirements of the mitochondrial heavy-strand promoters.Transcription · 2017Article
- Mitochondrial transcription in mammalian cells.Frontiers in bioscience (Landmark edition) · 2017Review
- Therapeutic efficacy of the small molecule GS-5734 against Ebola virus in rhesus monkeys.Nature · 2016Article
- Expression and Purification of Mitochondrial RNA Polymerase and Transcription Factor A from Drosophila melanogaster.Methods in molecular biology (Clifton, N.J.) · 2016Article
- Inhibition of hepatitis C virus replication by GS-6620, a potent C-nucleoside monophosphate prodrug.Antimicrobial agents and chemotherapy · 2014Article
- Accessorizing the human mitochondrial transcription machinery.Trends in biochemical sciences · 2013Review
- Human Cockayne syndrome B protein reciprocally communicates with mitochondrial proteins and promotes transcriptional elongation.Nucleic acids research · 2012Article
- Conservation of promoter melting mechanisms in divergent regions of the single-subunit RNA polymerases.Biochemistry · 2012Article
- Transcription from the second heavy-strand promoter of human mtDNA is repressed by transcription factor A in vitro.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- Transcriptional requirements of the distal heavy-strand promoter of mtDNA.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- Transcriptional activation by mitochondrial transcription factor A involves preferential distortion of promoter DNA.Nucleic acids research · 2012Article
- Sensitivity of mitochondrial transcription and resistance of RNA polymerase II dependent nuclear transcription to antiviral ribonucleosides.PLoS pathogens · 2012Article
- Mitochondrial ribosomal protein L12 selectively associates with human mitochondrial RNA polymerase to activate transcription.Proceedings of the National Academy of Sciences of the United States of America · 2011Article
- The mitochondrial transcription and packaging factor Tfam imposes a U-turn on mitochondrial DNA.Nature structural & molecular biology · 2011Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We have reconstituted human mitochondrial transcription in vitro on DNA oligonucleotide templates representing the light strand and heavy strand-1 promoters using protein components (RNA polymerase and transcription factors A and B2) isolated from Escherichia coli. We show that 1 eq of each transcription factor and polymerase relative to the promoter is required to assemble a functional initiation complex. The light strand promoter is at least 2-fold more efficient than the heavy strand-1 promoter, but this difference cannot be explained solely by the differences in the interaction of the transcription machinery with the different promoters. In both cases, the rate-limiting step for production of the first phosphodiester bond is open complex formation. Open complex formation requires both transcription factors; however, steps immediately thereafter only require transcription factor B2. The concentration of nucleotide required for production of the first dinucleotide product is substantially higher than that required for subsequent cycles of nucleotide addition. In vitro, promoter-specific differences in post-initiation control of transcription exist, as well as a second rate-limiting step that controls conversion of the transcription initiation complex into a transcription elongation complex. Rate-limiting steps of the biochemical pathways are often those that are targeted for regulation. Like the more complex multisubunit transcription systems, multiple steps may exist for control of transcription in human mitochondria. The tools and mechanistic framework presented here will facilitate not only the discovery of mechanisms regulating human mitochondrial transcription but also interrogation of the structure, function, and mechanism of the complexes that are regulated during human mitochondrial transcription.
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