Pre-steady state kinetic studies show that an abasic site is a cognate lesion for the yeast Rev1 protein

John M Pryor; M Todd Washington

doi:10.1016/j.dnarep.2011.08.011

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Pre-steady state kinetic studies show that an abasic site is a cognate lesion for the yeast Rev1 protein

Journal article

Peer reviewed

Pre-steady state kinetic studies show that an abasic site is a cognate lesion for the yeast Rev1 protein

John M Pryor and M Todd Washington

DNA repair, Vol.10(11), pp.1138-1144

11/10/2011

DOI: 10.1016/j.dnarep.2011.08.011

PMCID: PMC3197757

PMID: 21975119

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Abstract

Rev1 is a eukaryotic DNA polymerase that rescues replication forks stalled at sites of DNA damage by inserting nucleotides opposite the damaged template bases. Yeast genetic studies suggest that Rev1 plays an important role in rescuing replication forks stalled at one of the most common forms of DNA damage, an abasic site; however, steady state kinetic studies suggest that an abasic site acts as a significant block to nucleotide incorporation by Rev1. Here we examined the pre-steady state kinetics of nucleotide incorporation by yeast Rev1 with damaged and non-damaged DNA substrates. We found that yeast Rev1 is capable of rapid nucleotide incorporation, but only a small fraction of the protein molecules possessed this robust activity. We characterized the nucleotide incorporation by the catalytically robust fraction of yeast Rev1 and found that it efficiently incorporated dCTP opposite a template abasic site under pre-steady state conditions. We conclude from these studies that the abasic site is a cognate lesion for Rev1.

Yeasts - genetics

Catalytic Domain

Substrate Specificity

Deoxycytosine Nucleotides - metabolism

DNA - metabolism

DNA-Directed DNA Polymerase - chemistry

DNA-Directed DNA Polymerase - metabolism

Kinetics

Yeasts - enzymology

Details

Title: Subtitle: Pre-steady state kinetic studies show that an abasic site is a cognate lesion for the yeast Rev1 protein
Creators: John M Pryor - Department of Biochemistry, University of Iowa College of Medicine, Iowa City, IA 52242-1109, United States
M Todd Washington
Resource Type: Journal article
Publication Details: DNA repair, Vol.10(11), pp.1138-1144
DOI: 10.1016/j.dnarep.2011.08.011
PMID: 21975119
PMCID: PMC3197757
NLM abbreviation: DNA Repair (Amst)
ISSN: 1568-7864
eISSN: 1568-7856
Publisher: Netherlands
Grant note: GM081433 / NIGMS NIH HHS R01 GM081433 / NIGMS NIH HHS R01 GM081433-03 / NIGMS NIH HHS
Language: English
Date published: 11/10/2011
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984025299302771

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