HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 282, Issue 4, 2324-2332, January 26, 2007

This Article Full Text Full Text (PDF) All Versions of this Article: 282/4/2324    most recent M609591200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by McElhinny, S. A. N. Articles by Kunkel, T. A. Search for Related Content PubMed PubMed Citation Articles by McElhinny, S. A. N. Articles by Kunkel, T. A. Social Bookmarking                      What's this?

Inefficient Proofreading and Biased Error Rates during Inaccurate DNA Synthesis by a Mutant Derivative of Saccharomyces cerevisiae DNA Polymerase ^*

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

From the Laboratory of Molecular Genetics and Laboratory of Structural Biology, NIEHS, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, North Carolina 27709 and the Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110

DNA polymerase (pol ) is a high fidelity eukaryotic enzyme that participates in DNA repair and is essential for DNA replication. Toward the goal of dissecting its multiple biological functions, here we describe the biochemical properties of Saccharomyces cerevisiae pol with a methionine replacing conserved leucine 612 at the polymerase active site. Compared with wild type pol , L612M pol has normal processivity and slightly higher polymerase specific activity. L612M pol also has normal 3' exonuclease activity, yet it is impaired in partitioning mismatches to the exonuclease active site, thereby reducing DNA synthesis fidelity. Error rates in vitro for L612M pol are elevated for both base substitutions and single base deletions but in a highly biased manner. For each of the six possible pairs of reciprocal mismatches that could arise during replication of complementary DNA strands to account for any particular base substitution in vivo (e.g. T-dGMP or A-dCMP for T to C transitions), L612M pol error rates are substantially higher for one mismatch than the other. These results provide a biochemical explanation for our observation, which confirms earlier genetic studies, that a haploid pol3-L612M S. cerevisiae strain has an elevated spontaneous mutation rate that is likely due to reduced replication fidelity in vivo.

Received for publication, October 11, 2006 , and in revised form, November 13, 2006.

^* This work was funded in part by the Intramural Research Program of the NIEHS, National Institutes of Health (to T. A. K.) and in part by National Institutes of Health Grant GM32431 (to P. M. J. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed. Tel.: 919-541-2644; Fax: 919-541-7613; E-mail: kunkel{at}niehs.nih.gov.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. Suzuki, A. Niimi, S. Limsirichaikul, S. Tomida, Q. Miao Huang, S. Izuta, J. Usukura, Y. Itoh, T. Hishida, T. Akashi, et al. PCNA Mono-Ubiquitination and Activation of Translesion DNA Polymerases by DNA Polymerase {alpha} J. Biochem., July 1, 2009; 146(1): 13 - 21. [Abstract] [Full Text] [PDF]

				J. E. Stone, G. E. Kissling, S. A. Lujan, I. B. Rogozin, C. M. Stith, P. M. J. Burgers, and T. A. Kunkel Low-fidelity DNA synthesis by the L979F mutator derivative of Saccharomyces cerevisiae DNA polymerase {zeta} Nucleic Acids Res., June 1, 2009; 37(11): 3774 - 3787. [Abstract] [Full Text] [PDF]

				X. Zhong, L. C. Pedersen, and T. A. Kunkel Characterization of a replicative DNA polymerase mutant with reduced fidelity and increased translesion synthesis capacity Nucleic Acids Res., July 1, 2008; 36(12): 3892 - 3904. [Abstract] [Full Text] [PDF]

				R. N. Venkatesan, P. M. Treuting, E. D. Fuller, R. E. Goldsby, T. H. Norwood, T. A. Gooley, W. C. Ladiges, B. D. Preston, and L. A. Loeb Mutation at the Polymerase Active Site of Mouse DNA Polymerase {delta} Increases Genomic Instability and Accelerates Tumorigenesis Mol. Cell. Biol., November 1, 2007; 27(21): 7669 - 7682. [Abstract] [Full Text] [PDF]

				R. L. Eoff, A. Irimia, K. C. Angel, M. Egli, and F. P. Guengerich Hydrogen Bonding of 7,8-Dihydro-8-oxodeoxyguanosine with a Charged Residue in the Little Finger Domain Determines Miscoding Events in Sulfolobus solfataricus DNA Polymerase Dpo4 J. Biol. Chem., July 6, 2007; 282(27): 19831 - 19843. [Abstract] [Full Text] [PDF]