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A novel intermediate in transcription initiation by human mitochondrial RNA polymerase
The mitochondrial genome is transcribed by a single-subunit T7 phage-like RNA polymerase (mtRNAP), structurally unrelated to cellular RNAPs. In higher eukaryotes, mtRNAP requires two transcription factors for efficient initiation—TFAM, a major nucleoid protein, and TFB2M, a transient component of mt...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3973326/ https://www.ncbi.nlm.nih.gov/pubmed/24393772 http://dx.doi.org/10.1093/nar/gkt1356 |
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author | Morozov, Yaroslav I. Agaronyan, Karen Cheung, Alan C. M. Anikin, Michael Cramer, Patrick Temiakov, Dmitry |
author_facet | Morozov, Yaroslav I. Agaronyan, Karen Cheung, Alan C. M. Anikin, Michael Cramer, Patrick Temiakov, Dmitry |
author_sort | Morozov, Yaroslav I. |
collection | PubMed |
description | The mitochondrial genome is transcribed by a single-subunit T7 phage-like RNA polymerase (mtRNAP), structurally unrelated to cellular RNAPs. In higher eukaryotes, mtRNAP requires two transcription factors for efficient initiation—TFAM, a major nucleoid protein, and TFB2M, a transient component of mtRNAP catalytic site. The mechanisms behind assembly of the mitochondrial transcription machinery and its regulation are poorly understood. We isolated and identified a previously unknown human mitochondrial transcription intermediate—a pre-initiation complex that includes mtRNAP, TFAM and promoter DNA. Using protein–protein cross-linking, we demonstrate that human TFAM binds to the N-terminal domain of mtRNAP, which results in bending of the promoter DNA around mtRNAP. The subsequent recruitment of TFB2M induces promoter melting and formation of an open initiation complex. Our data indicate that the pre-initiation complex is likely to be an important target for transcription regulation and provide basis for further structural, biochemical and biophysical studies of mitochondrial transcription. |
format | Online Article Text |
id | pubmed-3973326 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-39733262014-04-04 A novel intermediate in transcription initiation by human mitochondrial RNA polymerase Morozov, Yaroslav I. Agaronyan, Karen Cheung, Alan C. M. Anikin, Michael Cramer, Patrick Temiakov, Dmitry Nucleic Acids Res Nucleic Acid Enzymes The mitochondrial genome is transcribed by a single-subunit T7 phage-like RNA polymerase (mtRNAP), structurally unrelated to cellular RNAPs. In higher eukaryotes, mtRNAP requires two transcription factors for efficient initiation—TFAM, a major nucleoid protein, and TFB2M, a transient component of mtRNAP catalytic site. The mechanisms behind assembly of the mitochondrial transcription machinery and its regulation are poorly understood. We isolated and identified a previously unknown human mitochondrial transcription intermediate—a pre-initiation complex that includes mtRNAP, TFAM and promoter DNA. Using protein–protein cross-linking, we demonstrate that human TFAM binds to the N-terminal domain of mtRNAP, which results in bending of the promoter DNA around mtRNAP. The subsequent recruitment of TFB2M induces promoter melting and formation of an open initiation complex. Our data indicate that the pre-initiation complex is likely to be an important target for transcription regulation and provide basis for further structural, biochemical and biophysical studies of mitochondrial transcription. Oxford University Press 2014-04 2014-01-06 /pmc/articles/PMC3973326/ /pubmed/24393772 http://dx.doi.org/10.1093/nar/gkt1356 Text en © The Author(s) 2014. Published by Oxford University Press. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Nucleic Acid Enzymes Morozov, Yaroslav I. Agaronyan, Karen Cheung, Alan C. M. Anikin, Michael Cramer, Patrick Temiakov, Dmitry A novel intermediate in transcription initiation by human mitochondrial RNA polymerase |
title | A novel intermediate in transcription initiation by human mitochondrial RNA polymerase |
title_full | A novel intermediate in transcription initiation by human mitochondrial RNA polymerase |
title_fullStr | A novel intermediate in transcription initiation by human mitochondrial RNA polymerase |
title_full_unstemmed | A novel intermediate in transcription initiation by human mitochondrial RNA polymerase |
title_short | A novel intermediate in transcription initiation by human mitochondrial RNA polymerase |
title_sort | novel intermediate in transcription initiation by human mitochondrial rna polymerase |
topic | Nucleic Acid Enzymes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3973326/ https://www.ncbi.nlm.nih.gov/pubmed/24393772 http://dx.doi.org/10.1093/nar/gkt1356 |
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