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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...

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Autores principales: Kondratick, Christine M., Boehm, Elizabeth M., Dieckman, Lynne M., Powers, Kyle T., Sanchez, Julio C., Mueting, Samuel R., Washington, M. Todd
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
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.
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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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