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Break-Induced Replication Is Highly Inaccurate
DNA must be synthesized for purposes of genome duplication and DNA repair. While the former is a highly accurate process, short-patch synthesis associated with repair of DNA damage is often error-prone. Break-induced replication (BIR) is a unique cellular process that mimics normal DNA replication i...
Autores principales: | , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3039667/ https://www.ncbi.nlm.nih.gov/pubmed/21347245 http://dx.doi.org/10.1371/journal.pbio.1000594 |
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author | Deem, Angela Keszthelyi, Andrea Blackgrove, Tiffany Vayl, Alexandra Coffey, Barbara Mathur, Ruchi Chabes, Andrei Malkova, Anna |
author_facet | Deem, Angela Keszthelyi, Andrea Blackgrove, Tiffany Vayl, Alexandra Coffey, Barbara Mathur, Ruchi Chabes, Andrei Malkova, Anna |
author_sort | Deem, Angela |
collection | PubMed |
description | DNA must be synthesized for purposes of genome duplication and DNA repair. While the former is a highly accurate process, short-patch synthesis associated with repair of DNA damage is often error-prone. Break-induced replication (BIR) is a unique cellular process that mimics normal DNA replication in its processivity, rate, and capacity to duplicate hundreds of kilobases, but is initiated at double-strand breaks (DSBs) rather than at replication origins. Here we employed a series of frameshift reporters to measure mutagenesis associated with BIR in Saccharomyces cerevisiae. We demonstrate that BIR DNA synthesis is intrinsically inaccurate over the entire path of the replication fork, as the rate of frameshift mutagenesis during BIR is up to 2,800-fold higher than during normal replication. Importantly, this high rate of mutagenesis was observed not only close to the DSB where BIR is less stable, but also far from the DSB where the BIR replication fork is fast and stabilized. We established that polymerase proofreading and mismatch repair correct BIR errors. Also, dNTP levels were elevated during BIR, and this contributed to BIR-related mutagenesis. We propose that a high level of DNA polymerase errors that is not fully compensated by error-correction mechanisms is largely responsible for mutagenesis during BIR, with Pol δ generating many of the mutagenic errors. We further postulate that activation of BIR in eukaryotic cells may significantly contribute to accumulation of mutations that fuel cancer and evolution. |
format | Text |
id | pubmed-3039667 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-30396672011-02-23 Break-Induced Replication Is Highly Inaccurate Deem, Angela Keszthelyi, Andrea Blackgrove, Tiffany Vayl, Alexandra Coffey, Barbara Mathur, Ruchi Chabes, Andrei Malkova, Anna PLoS Biol Research Article DNA must be synthesized for purposes of genome duplication and DNA repair. While the former is a highly accurate process, short-patch synthesis associated with repair of DNA damage is often error-prone. Break-induced replication (BIR) is a unique cellular process that mimics normal DNA replication in its processivity, rate, and capacity to duplicate hundreds of kilobases, but is initiated at double-strand breaks (DSBs) rather than at replication origins. Here we employed a series of frameshift reporters to measure mutagenesis associated with BIR in Saccharomyces cerevisiae. We demonstrate that BIR DNA synthesis is intrinsically inaccurate over the entire path of the replication fork, as the rate of frameshift mutagenesis during BIR is up to 2,800-fold higher than during normal replication. Importantly, this high rate of mutagenesis was observed not only close to the DSB where BIR is less stable, but also far from the DSB where the BIR replication fork is fast and stabilized. We established that polymerase proofreading and mismatch repair correct BIR errors. Also, dNTP levels were elevated during BIR, and this contributed to BIR-related mutagenesis. We propose that a high level of DNA polymerase errors that is not fully compensated by error-correction mechanisms is largely responsible for mutagenesis during BIR, with Pol δ generating many of the mutagenic errors. We further postulate that activation of BIR in eukaryotic cells may significantly contribute to accumulation of mutations that fuel cancer and evolution. Public Library of Science 2011-02-15 /pmc/articles/PMC3039667/ /pubmed/21347245 http://dx.doi.org/10.1371/journal.pbio.1000594 Text en Deem et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Deem, Angela Keszthelyi, Andrea Blackgrove, Tiffany Vayl, Alexandra Coffey, Barbara Mathur, Ruchi Chabes, Andrei Malkova, Anna Break-Induced Replication Is Highly Inaccurate |
title | Break-Induced Replication Is Highly Inaccurate |
title_full | Break-Induced Replication Is Highly Inaccurate |
title_fullStr | Break-Induced Replication Is Highly Inaccurate |
title_full_unstemmed | Break-Induced Replication Is Highly Inaccurate |
title_short | Break-Induced Replication Is Highly Inaccurate |
title_sort | break-induced replication is highly inaccurate |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3039667/ https://www.ncbi.nlm.nih.gov/pubmed/21347245 http://dx.doi.org/10.1371/journal.pbio.1000594 |
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