Substitution of a residue contacting the triphosphate moiety of the incoming nucleotide increases the fidelity of yeast DNA polymerase ζ

Craig A Howell; Christine M Kondratick; M. Todd Washington

doi:10.1093/nar/gkn023

Back

Substitution of a residue contacting the triphosphate moiety of the incoming nucleotide increases the fidelity of yeast DNA polymerase ζ

Journal article

Open access

Peer reviewed

Substitution of a residue contacting the triphosphate moiety of the incoming nucleotide increases the fidelity of yeast DNA polymerase ζ

Craig A Howell, Christine M Kondratick and M. Todd Washington

Nucleic acids research, Vol.36(5), pp.1731-1740

03/2008

DOI: 10.1093/nar/gkn023

PMCID: PMC2275142

PMID: 18263611

Files and links (1)

url

https://doi.org/10.1093/nar/gkn023View

Published (Version of record) Open Access

Abstract

DNA polymerase ζ (pol ζ), which is required for DNA damage-induced mutagenesis, functions in the error-prone replication of a wide range of DNA lesions. During this process, pol ζ extends from nucleotides incorporated opposite template lesions by other polymerases. Unlike classical polymerases, pol ζ efficiently extends from primer-terminal base pairs containing mismatches or lesions, and it synthesizes DNA with moderate fidelity. Here we describe genetic and biochemical studies of three yeast pol ζ mutant proteins containing substitutions of highly conserved amino acid residues that contact the triphosphate moiety of the incoming nucleotide. The R1057A and K1086A proteins do not complement the rev3Δ mutation, and these proteins have significantly reduced polymerase activity relative to the wild-type protein. In contrast, the K1061A protein partially complements the rev3Δ mutation and has nearly normal polymerase activity. Interestingly, the K1061A protein has increased fidelity relative to wild-type pol ζ and is somewhat less efficient at extending from mismatched primer-terminal base pairs. These findings have important implications both for the evolutionary divergence of pol ζ from classical polymerases and for the mechanism by which this enzyme accommodates distortions in the DNA caused by mismatches and lesions.

Details

Title: Subtitle: Substitution of a residue contacting the triphosphate moiety of the incoming nucleotide increases the fidelity of yeast DNA polymerase ζ
Creators: Craig A Howell
Christine M Kondratick
M. Todd Washington
Resource Type: Journal article
Publication Details: Nucleic acids research, Vol.36(5), pp.1731-1740
DOI: 10.1093/nar/gkn023
PMID: 18263611
PMCID: PMC2275142
NLM abbreviation: Nucleic Acids Res
ISSN: 0305-1048
eISSN: 1362-4962
Publisher: Oxford University Press
Language: English
Date published: 03/2008
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984024509602771

Metrics

18 Record Views

7 Times Cited - Web of Science