Division of Labor at the Eukaryotic Replication Fork

Stephanie A. Nick McElhinny; Dmitry A. Gordenin; Carrie M. Stith; Peter M.J. Burgers; Thomas A. Kunkel

doi:10.1016/j.molcel.2008.02.022

Division of Labor at the Eukaryotic Replication Fork

Stephanie A. Nick McElhinny, Dmitry A. Gordenin, Carrie M. Stith, Peter M.J. Burgers, Thomas A. Kunkel

Research output: Contribution to journal › Article › peer-review

389 Scopus citations

Abstract

DNA polymerase δ (Pol δ) and DNA polymerase ε (Pol ε) are both required for efficient replication of the nuclear genome, yet the division of labor between these enzymes has remained unclear for many years. Here we investigate the contribution of Pol δ to replication of the leading and lagging strand templates in Saccharomyces cerevisiae using a mutant Pol δ allele (pol3-L612M) whose error rate is higher for one mismatch (e.g., T•dGTP) than for its complement (A•dCTP). We find that strand-specific mutation rates strongly depend on the orientation of a reporter gene relative to an adjacent replication origin, in a manner implying that >90% of Pol δ replication is performed using the lagging strand template. When combined with recent evidence implicating Pol ε in leading strand replication, these data support a model of the replication fork wherein the leading and lagging strand templates are primarily copied by Pol ε and Pol δ, respectively.

Original language	English
Pages (from-to)	137-144
Number of pages	8
Journal	Molecular cell
Volume	30
Issue number	2
DOIs	https://doi.org/10.1016/j.molcel.2008.02.022
State	Published - Apr 25 2008

Keywords

CELLCYCLE
DNA

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10.1016/j.molcel.2008.02.022

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title = "Division of Labor at the Eukaryotic Replication Fork",

abstract = "DNA polymerase δ (Pol δ) and DNA polymerase ε (Pol ε) are both required for efficient replication of the nuclear genome, yet the division of labor between these enzymes has remained unclear for many years. Here we investigate the contribution of Pol δ to replication of the leading and lagging strand templates in Saccharomyces cerevisiae using a mutant Pol δ allele (pol3-L612M) whose error rate is higher for one mismatch (e.g., T•dGTP) than for its complement (A•dCTP). We find that strand-specific mutation rates strongly depend on the orientation of a reporter gene relative to an adjacent replication origin, in a manner implying that >90% of Pol δ replication is performed using the lagging strand template. When combined with recent evidence implicating Pol ε in leading strand replication, these data support a model of the replication fork wherein the leading and lagging strand templates are primarily copied by Pol ε and Pol δ, respectively.",

keywords = "CELLCYCLE, DNA",

author = "{Nick McElhinny}, {Stephanie A.} and Gordenin, {Dmitry A.} and Stith, {Carrie M.} and Burgers, {Peter M.J.} and Kunkel, {Thomas A.}",

note = "Funding Information: We are particularly grateful to Dr. Youri Pavlov (University of Nebraska Medical Center) for discussions and advice during the early stages of this work. We sincerely thank Dr. John Drake (NIEHS), Dr. Roel Schaaper (NIEHS), and Dr. Dale Ramsden (University of North Carolina-Chapel Hill) for critical reading of and thoughtful comments on the manuscript; Dr. Robert Kokoska for help in strain construction; and the NIEHS DNA Sequencing Core Laboratory for expert assistance in sequence analysis of mutant clones. This work was funded in part by the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences to T.A.K., and in part by National Institutes of Health Grant GM32431 to P.M.J.B. ",

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AU - Gordenin, Dmitry A.

AU - Stith, Carrie M.

AU - Burgers, Peter M.J.

AU - Kunkel, Thomas A.

N1 - Funding Information: We are particularly grateful to Dr. Youri Pavlov (University of Nebraska Medical Center) for discussions and advice during the early stages of this work. We sincerely thank Dr. John Drake (NIEHS), Dr. Roel Schaaper (NIEHS), and Dr. Dale Ramsden (University of North Carolina-Chapel Hill) for critical reading of and thoughtful comments on the manuscript; Dr. Robert Kokoska for help in strain construction; and the NIEHS DNA Sequencing Core Laboratory for expert assistance in sequence analysis of mutant clones. This work was funded in part by the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences to T.A.K., and in part by National Institutes of Health Grant GM32431 to P.M.J.B.

PY - 2008/4/25

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N2 - DNA polymerase δ (Pol δ) and DNA polymerase ε (Pol ε) are both required for efficient replication of the nuclear genome, yet the division of labor between these enzymes has remained unclear for many years. Here we investigate the contribution of Pol δ to replication of the leading and lagging strand templates in Saccharomyces cerevisiae using a mutant Pol δ allele (pol3-L612M) whose error rate is higher for one mismatch (e.g., T•dGTP) than for its complement (A•dCTP). We find that strand-specific mutation rates strongly depend on the orientation of a reporter gene relative to an adjacent replication origin, in a manner implying that >90% of Pol δ replication is performed using the lagging strand template. When combined with recent evidence implicating Pol ε in leading strand replication, these data support a model of the replication fork wherein the leading and lagging strand templates are primarily copied by Pol ε and Pol δ, respectively.

AB - DNA polymerase δ (Pol δ) and DNA polymerase ε (Pol ε) are both required for efficient replication of the nuclear genome, yet the division of labor between these enzymes has remained unclear for many years. Here we investigate the contribution of Pol δ to replication of the leading and lagging strand templates in Saccharomyces cerevisiae using a mutant Pol δ allele (pol3-L612M) whose error rate is higher for one mismatch (e.g., T•dGTP) than for its complement (A•dCTP). We find that strand-specific mutation rates strongly depend on the orientation of a reporter gene relative to an adjacent replication origin, in a manner implying that >90% of Pol δ replication is performed using the lagging strand template. When combined with recent evidence implicating Pol ε in leading strand replication, these data support a model of the replication fork wherein the leading and lagging strand templates are primarily copied by Pol ε and Pol δ, respectively.

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Division of Labor at the Eukaryotic Replication Fork

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