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Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis
Proliferating cell nuclear antigen (PCNA) plays an essential role in DNA replication and repair by interacting with a large number of proteins involved in these processes. Two amino acid substitutions in PCNA, both located at the subunit interface, have previously been shown to block translesion syn...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4892588/ https://www.ncbi.nlm.nih.gov/pubmed/27258147 http://dx.doi.org/10.1371/journal.pone.0157023 |
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author | Kondratick, Christine M. Boehm, Elizabeth M. Dieckman, Lynne M. Powers, Kyle T. Sanchez, Julio C. Mueting, Samuel R. Washington, M. Todd |
author_facet | Kondratick, Christine M. Boehm, Elizabeth M. Dieckman, Lynne M. Powers, Kyle T. Sanchez, Julio C. Mueting, Samuel R. Washington, M. Todd |
author_sort | Kondratick, Christine M. |
collection | PubMed |
description | Proliferating cell nuclear antigen (PCNA) plays an essential role in DNA replication and repair by interacting with a large number of proteins involved in these processes. Two amino acid substitutions in PCNA, both located at the subunit interface, have previously been shown to block translesion synthesis (TLS), a pathway for bypassing DNA damage during replication. To better understand the role of the subunit interface in TLS, we used random mutagenesis to generate a set of 33 PCNA mutants with substitutions at the subunit interface. We assayed the full set of mutants for viability and sensitivity to ultraviolet (UV) radiation. We then selected a subset of 17 mutants and measured their rates of cell growth, spontaneous mutagenesis, and UV-induced mutagenesis. All except three of these 17 mutants were partially or completely defective in induced mutagenesis, which indicates a partial or complete loss of TLS. These results demonstrate that the integrity of the subunit interface of PCNA is essential for efficient TLS and that even conservative substitutions have the potential to disrupt this process. |
format | Online Article Text |
id | pubmed-4892588 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-48925882016-06-16 Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis Kondratick, Christine M. Boehm, Elizabeth M. Dieckman, Lynne M. Powers, Kyle T. Sanchez, Julio C. Mueting, Samuel R. Washington, M. Todd PLoS One Research Article Proliferating cell nuclear antigen (PCNA) plays an essential role in DNA replication and repair by interacting with a large number of proteins involved in these processes. Two amino acid substitutions in PCNA, both located at the subunit interface, have previously been shown to block translesion synthesis (TLS), a pathway for bypassing DNA damage during replication. To better understand the role of the subunit interface in TLS, we used random mutagenesis to generate a set of 33 PCNA mutants with substitutions at the subunit interface. We assayed the full set of mutants for viability and sensitivity to ultraviolet (UV) radiation. We then selected a subset of 17 mutants and measured their rates of cell growth, spontaneous mutagenesis, and UV-induced mutagenesis. All except three of these 17 mutants were partially or completely defective in induced mutagenesis, which indicates a partial or complete loss of TLS. These results demonstrate that the integrity of the subunit interface of PCNA is essential for efficient TLS and that even conservative substitutions have the potential to disrupt this process. Public Library of Science 2016-06-03 /pmc/articles/PMC4892588/ /pubmed/27258147 http://dx.doi.org/10.1371/journal.pone.0157023 Text en © 2016 Kondratick 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 (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Kondratick, Christine M. Boehm, Elizabeth M. Dieckman, Lynne M. Powers, Kyle T. Sanchez, Julio C. Mueting, Samuel R. Washington, M. Todd Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis |
title | Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis |
title_full | Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis |
title_fullStr | Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis |
title_full_unstemmed | Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis |
title_short | Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis |
title_sort | identification of new mutations at the pcna subunit interface that block translesion synthesis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4892588/ https://www.ncbi.nlm.nih.gov/pubmed/27258147 http://dx.doi.org/10.1371/journal.pone.0157023 |
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