Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis

Christine M Kondratick; Elizabeth M Boehm; Lynne M Dieckman; Kyle T Powers; Julio C Sanchez; Samuel R Mueting; M Todd Washington

doi:10.1371/journal.pone.0157023

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Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis

Journal article

Open access

Peer reviewed

Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis

Christine M Kondratick, Elizabeth M Boehm, Lynne M Dieckman, Kyle T Powers, Julio C Sanchez, Samuel R Mueting and M Todd Washington

PloS one, Vol.11(6), pp.e0157023-e0157023

2016

DOI: 10.1371/journal.pone.0157023

PMCID: PMC4892588

PMID: 27258147

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Abstract

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.

Protein Structure, Secondary

Saccharomyces cerevisiae - genetics

Saccharomyces cerevisiae Proteins - genetics

Mutation - genetics

Proliferating Cell Nuclear Antigen - chemistry

Saccharomyces cerevisiae - metabolism

Proliferating Cell Nuclear Antigen - genetics

Ultraviolet Rays

Mutagenesis

Saccharomyces cerevisiae Proteins - metabolism

DNA Damage - genetics

DNA Replication - genetics

Protein Binding

Proliferating Cell Nuclear Antigen - metabolism

Saccharomyces cerevisiae Proteins - chemistry

Details

Title: Subtitle: Identification of New Mutations at the PCNA Subunit Interface that Block Translesion Synthesis
Creators: Christine M Kondratick - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, Iowa, 52242, United States of America
Elizabeth M Boehm - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, Iowa, 52242, United States of America
Lynne M Dieckman - Department of Chemistry, Creighton University, Omaha, Nebraska, 68178, United States of America
Kyle T Powers - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, Iowa, 52242, United States of America
Julio C Sanchez - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, Iowa, 52242, United States of America
Samuel R Mueting - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, Iowa, 52242, United States of America
M Todd Washington - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, Iowa, 52242, United States of America
Resource Type: Journal article
Publication Details: PloS one, Vol.11(6), pp.e0157023-e0157023
DOI: 10.1371/journal.pone.0157023
PMID: 27258147
PMCID: PMC4892588
NLM abbreviation: PLoS One
ISSN: 1932-6203
eISSN: 1932-6203
Publisher: Public Library of Science; United States
Grant note: R01 GM081433 / NIGMS NIH HHS P30 CA086862 / NCI NIH HHS R01 GM108027 / NIGMS NIH HHS
Language: English
Date published: 2016
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984024407102771

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