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Dpb2p, a Noncatalytic Subunit of DNA Polymerase ε, Contributes to the Fidelity of DNA Replication in Saccharomyces cerevisiae

Jaszczur, Malgorzata and Flis, Krzysztof and Rudzka, Justyna and Kraszewska, Joanna and Budd, Martin E. and Polaczek, Piotr and Campbell, Judith L. and Jonczyk, Piotr and Fijalkowska, Iwona J. (2008) Dpb2p, a Noncatalytic Subunit of DNA Polymerase ε, Contributes to the Fidelity of DNA Replication in Saccharomyces cerevisiae. Genetics, 178 (2). pp. 633-647. ISSN 0016-6731. PMCID PMC2248333. doi:10.1534/genetics.107.082818.

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Most replicases are multi-subunit complexes. DNA polymerase epsilon from Saccharomyces cerevisiae is composed of four subunits: Pol2p, Dpb2p, Dpb3p, and Dpb4p. Pol2p and Dpb2p are essential. To investigate a possible role for the Dpb2p subunit in maintaining the fidelity of DNA replication, we isolated temperature-sensitive mutants in the DPB2 gene. Several of the newly isolated dpb2 alleles are strong mutators, exhibiting mutation rates equivalent to pol2 mutants defective in the 3′ → 5′ proofreading exonuclease (pol2-4) or to mutants defective in mismatch repair (msh6). The dpb2 pol2-4 and dpb2 msh6 double mutants show a synergistic increase in mutation rate, indicating that the mutations arising in the dpb2 mutants are due to DNA replication errors normally corrected by mismatch repair. The dpb2 mutations decrease the affinity of Dpb2p for the Pol2p subunit as measured by two-hybrid analysis, providing a possible mechanistic explanation for the loss of high-fidelity synthesis. Our results show that DNA polymerase subunits other than those housing the DNA polymerase and 3′ → 5′ exonuclease are essential in controlling the level of spontaneous mutagenesis and genetic stability in yeast cells.

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Additional Information:© 2008 by the Genetics Society of America. Manuscript received October 3, 2007; Accepted for publication November 19, 2007. We thank Katarzyna Bebenek of the National Institute of Environmental Health Sciences (Research Triangle Park) and Zygmunt Ciesla of the Institute of Biochemistry and Biophysics Polish Academy of Sciences (Warsaw) for their critical reading of the manuscript; Anna Gajda for her excellent technical assistance; Youri Pavlov of the Eppley Institute for Research in Cancer, University of Nebraska Medical Center (Omaha); and Hiroyuki Araki, Division of Microbial Genetics, National Institute of Genetics, Research Organization of Information and Systems (Shizuoka, Japan) for providing S. cerevisiae strains. We thank also T. Mason (Department of Biochemistry, University of Massachusetts, Amherst, MA) for anti-Hts1p antibody. This work was supported by grant 2P04A05126 from the Polish Ministry of Science and Higher Education to M.J., K.F., P.J., and I.J.F; and by U. S. Public Health Service grants TW006463 [Fogarty International Collaboration Award (FIRCA)] to I.J.F., P.J., and J.L.C. and GM25508 to J.L.C.
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Ministry of Science and Higher Education (Poland)2P04A05126
Issue or Number:2
PubMed Central ID:PMC2248333
Record Number:CaltechAUTHORS:20200603-153936657
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Official Citation:Dpb2p, a Noncatalytic Subunit of DNA Polymerase ε, Contributes to the Fidelity of DNA Replication in Saccharomyces cerevisiae. Malgorzata Jaszczur, Krzysztof Flis, Justyna Rudzka, Joanna Kraszewska, Martin E. Budd, Piotr Polaczek, Judith L. Campbell, Piotr Jonczyk and Iwona J. Fijalkowska. GENETICS. February 1, 2008 vol. 178 no. 2 633-647;
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:103681
Deposited By: Tony Diaz
Deposited On:03 Jun 2020 22:59
Last Modified:16 Nov 2021 18:23

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