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Error-Prone Translesion DNA Synthesis by Escherichia coli DNA Polymerase IV (DinB) on Templates Containing 1,2-dihydro-2-oxoadenine
Escherichia coli DNA polymerase IV (Pol IV) is involved in bypass replication of damaged bases in DNA. Reactive oxygen species (ROS) are generated continuously during normal metabolism and as a result of exogenous stress such as ionizing radiation. ROS induce various kinds of base damage in DNA. It...
Autores principales: | , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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SAGE-Hindawi Access to Research
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2952919/ https://www.ncbi.nlm.nih.gov/pubmed/20976264 http://dx.doi.org/10.4061/2010/807579 |
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author | Hori, Masaki Yonekura, Shin-Ichiro Nohmi, Takehiko Gruz, Petr Sugiyama, Hiroshi Yonei, Shuji Zhang-Akiyama, Qiu-Mei |
author_facet | Hori, Masaki Yonekura, Shin-Ichiro Nohmi, Takehiko Gruz, Petr Sugiyama, Hiroshi Yonei, Shuji Zhang-Akiyama, Qiu-Mei |
author_sort | Hori, Masaki |
collection | PubMed |
description | Escherichia coli DNA polymerase IV (Pol IV) is involved in bypass replication of damaged bases in DNA. Reactive oxygen species (ROS) are generated continuously during normal metabolism and as a result of exogenous stress such as ionizing radiation. ROS induce various kinds of base damage in DNA. It is important to examine whether Pol IV is able to bypass oxidatively damaged bases. In this study, recombinant Pol IV was incubated with oligonucleotides containing thymine glycol (dTg), 5-formyluracil (5-fodU), 5-hydroxymethyluracil (5-hmdU), 7,8-dihydro-8-oxoguanine (8-oxodG) and 1,2-dihydro-2-oxoadenine (2-oxodA). Primer extension assays revealed that Pol IV preferred to insert dATP opposite 5-fodU and 5-hmdU, while it inefficiently inserted nucleotides opposite dTg. Pol IV inserted dCTP and dATP opposite 8-oxodG, while the ability was low. It inserted dCTP more effectively than dTTP opposite 2-oxodA. Pol IV's ability to bypass these lesions decreased in the order: 2-oxodA > 5-fodU~5-hmdU > 8-oxodG > dTg. The fact that Pol IV preferred to insert dCTP opposite 2-oxodA suggests the mutagenic potential of 2-oxodA leading to A:T→G:C transitions. Hydrogen peroxide caused an ~2-fold increase in A:T→G:C mutations in E. coli, while the increase was significantly greater in E. coli overexpressing Pol IV. These results indicate that Pol IV may be involved in ROS-enhanced A:T→G:C mutations. |
format | Text |
id | pubmed-2952919 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | SAGE-Hindawi Access to Research |
record_format | MEDLINE/PubMed |
spelling | pubmed-29529192010-10-25 Error-Prone Translesion DNA Synthesis by Escherichia coli DNA Polymerase IV (DinB) on Templates Containing 1,2-dihydro-2-oxoadenine Hori, Masaki Yonekura, Shin-Ichiro Nohmi, Takehiko Gruz, Petr Sugiyama, Hiroshi Yonei, Shuji Zhang-Akiyama, Qiu-Mei J Nucleic Acids Research Article Escherichia coli DNA polymerase IV (Pol IV) is involved in bypass replication of damaged bases in DNA. Reactive oxygen species (ROS) are generated continuously during normal metabolism and as a result of exogenous stress such as ionizing radiation. ROS induce various kinds of base damage in DNA. It is important to examine whether Pol IV is able to bypass oxidatively damaged bases. In this study, recombinant Pol IV was incubated with oligonucleotides containing thymine glycol (dTg), 5-formyluracil (5-fodU), 5-hydroxymethyluracil (5-hmdU), 7,8-dihydro-8-oxoguanine (8-oxodG) and 1,2-dihydro-2-oxoadenine (2-oxodA). Primer extension assays revealed that Pol IV preferred to insert dATP opposite 5-fodU and 5-hmdU, while it inefficiently inserted nucleotides opposite dTg. Pol IV inserted dCTP and dATP opposite 8-oxodG, while the ability was low. It inserted dCTP more effectively than dTTP opposite 2-oxodA. Pol IV's ability to bypass these lesions decreased in the order: 2-oxodA > 5-fodU~5-hmdU > 8-oxodG > dTg. The fact that Pol IV preferred to insert dCTP opposite 2-oxodA suggests the mutagenic potential of 2-oxodA leading to A:T→G:C transitions. Hydrogen peroxide caused an ~2-fold increase in A:T→G:C mutations in E. coli, while the increase was significantly greater in E. coli overexpressing Pol IV. These results indicate that Pol IV may be involved in ROS-enhanced A:T→G:C mutations. SAGE-Hindawi Access to Research 2010-09-26 /pmc/articles/PMC2952919/ /pubmed/20976264 http://dx.doi.org/10.4061/2010/807579 Text en Copyright © 2010 Masaki Hori et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Hori, Masaki Yonekura, Shin-Ichiro Nohmi, Takehiko Gruz, Petr Sugiyama, Hiroshi Yonei, Shuji Zhang-Akiyama, Qiu-Mei Error-Prone Translesion DNA Synthesis by Escherichia coli DNA Polymerase IV (DinB) on Templates Containing 1,2-dihydro-2-oxoadenine |
title | Error-Prone Translesion DNA Synthesis by Escherichia coli DNA Polymerase IV (DinB) on Templates Containing 1,2-dihydro-2-oxoadenine |
title_full | Error-Prone Translesion DNA Synthesis by Escherichia coli DNA Polymerase IV (DinB) on Templates Containing 1,2-dihydro-2-oxoadenine |
title_fullStr | Error-Prone Translesion DNA Synthesis by Escherichia coli DNA Polymerase IV (DinB) on Templates Containing 1,2-dihydro-2-oxoadenine |
title_full_unstemmed | Error-Prone Translesion DNA Synthesis by Escherichia coli DNA Polymerase IV (DinB) on Templates Containing 1,2-dihydro-2-oxoadenine |
title_short | Error-Prone Translesion DNA Synthesis by Escherichia coli DNA Polymerase IV (DinB) on Templates Containing 1,2-dihydro-2-oxoadenine |
title_sort | error-prone translesion dna synthesis by escherichia coli dna polymerase iv (dinb) on templates containing 1,2-dihydro-2-oxoadenine |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2952919/ https://www.ncbi.nlm.nih.gov/pubmed/20976264 http://dx.doi.org/10.4061/2010/807579 |
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