PCNA tool belts and polymerase bridges form during translesion synthesis

Elizabeth M Boehm; Maria Spies; M Todd Washington

doi:10.1093/nar/gkw563

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PCNA tool belts and polymerase bridges form during translesion synthesis

Journal article

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Peer reviewed

PCNA tool belts and polymerase bridges form during translesion synthesis

Elizabeth M Boehm, Maria Spies and M Todd Washington

Nucleic acids research, Vol.44(17), pp.8250-8260

09/30/2016

DOI: 10.1093/nar/gkw563

PMCID: PMC5041468

PMID: 27325737

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Abstract

Large multi-protein complexes play important roles in many biological processes, including DNA replication and repair, transcription, and signal transduction. One of the challenges in studying such complexes is to understand their mechanisms of assembly and disassembly and their architectures. Using single-molecule total internal reflection (TIRF) microscopy, we have examined the assembly and disassembly of the multi-protein complex that carries out translesion synthesis, the error-prone replication of damaged DNA. We show that the ternary complexes containing proliferating cell nuclear antigen (PCNA) and two non-classical DNA polymerases, Rev1 and DNA polymerase η, have two architectures: PCNA tool belts and Rev1 bridges. Moreover, these complexes are dynamic and their architectures can interconvert without dissociation. The formation of PCNA tool belts and Rev1 bridges and the ability of these complexes to change architectures are likely means of facilitating selection of the appropriate non-classical polymerase and polymerase-switching events.

Nucleotidyltransferases - chemistry

Fluorescence

Mutant Proteins - metabolism

DNA-Directed DNA Polymerase - chemistry

Amino Acid Motifs

Saccharomyces cerevisiae - metabolism

Multiprotein Complexes - metabolism

Saccharomyces cerevisiae Proteins - metabolism

Protein Binding

Protein Domains

DNA - biosynthesis

DNA-Directed DNA Polymerase - metabolism

Kinetics

Nucleotidyltransferases - metabolism

Proliferating Cell Nuclear Antigen - metabolism

Binding Sites

Amino Acid Substitution

Saccharomyces cerevisiae Proteins - chemistry

Details

Title: Subtitle: PCNA tool belts and polymerase bridges form during translesion synthesis
Creators: Elizabeth M Boehm - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Maria Spies - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
M Todd Washington - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA todd-washington@uiowa.edu
Resource Type: Journal article
Publication Details: Nucleic acids research, Vol.44(17), pp.8250-8260
Publisher: England
DOI: 10.1093/nar/gkw563
PMID: 27325737
PMCID: PMC5041468
ISSN: 0305-1048
eISSN: 1362-4962
Grant note: R01 GM108617 / NIGMS NIH HHS T32 GM067795 / NIGMS NIH HHS R01 GM108027 / NIGMS NIH HHS R01 GM081433 / NIGMS NIH HHS P30 CA086862 / NCI NIH HHS
Language: English
Date published: 09/30/2016
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984025295202771

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