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

J. Biol. Chem., Vol. 281, Issue 7, 4486-4494, February 17, 2006

This Article Full Text Full Text (PDF) All Versions of this Article: 281/7/4486    most recent M510245200v1 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 Venkatesan, R. N. Articles by Loeb, L. A. Search for Related Content PubMed PubMed Citation Articles by Venkatesan, R. N. Articles by Loeb, L. A. Social Bookmarking                      What's this?

Mutator Phenotypes Caused by Substitution at a Conserved Motif A Residue in Eukaryotic DNA Polymerase ^*

From the Department of Pathology, University of Washington, Seattle, Washington 98195-7705

Eukaryotic DNA polymerase (Pol) replicates chromosomal DNA and is also involved in DNA repair and genetic recombination. Motif A in Pol , containing the sequence DXXXLYPSI, includes a catalytically essential aspartic acid as well as other conserved residues of unknown function. Here, we used site-directed mutagenesis to create all 19 amino acid substitutions for the conserved Leu⁶¹² in Motif A of Saccharomyces cerevisiae Pol . We show that substitutions at Leu⁶¹² differentially affect viability, sensitivity to genotoxic agents, cell cycle progression, and replication fidelity. The eight viable mutants contained Ile, Val, Thr, Met, Phe, Lys, Asn, or Gly substitutions. Individual substitutions varied greatly in the nature and extent of attendant phenotypic deficiencies, exhibiting mutation rates that ranged from near wild type to a 37-fold increase. The L612M mutant exhibited a 7-fold elevation of mutation rate but essentially no detectable effects on other phenotypes monitored; the L612T mutant showed a nearly wild type mutation rate together with marked hypersensitivity to genotoxic agents; and the L612G and L612N strains exhibited relatively high mutation rates and severe deficits overall. We compare our results with those for homologous substitutions in prokaryotic and eukaryotic DNA polymerases and discuss the implications of our findings for the role of Leu⁶¹² in replication fidelity.

Received for publication, September 19, 2005 , and in revised form, December 8, 2005.

^* This work was supported by National Institutes of Health Grants R01 CA102029 (to L. A. L.), R01 ES 09927 and R01 CA98243 (to B. D. P.), and P01 AG01751. 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 Present address: Abbott Laboratories, Abbott Park, IL 60064.

² To whom correspondence should be addressed. Tel.: 206-543-6015; Fax: 206-543-3967; E-mail: laloeb{at}u.washington.edu.

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]

				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]

				Z. F. Pursell, I. Isoz, E.-B. Lundstrom, E. Johansson, and T. A. Kunkel Regulation of B family DNA polymerase fidelity by a conserved active site residue: characterization of M644W, M644L and M644F mutants of yeast DNA polymerase {varepsilon} Nucleic Acids Res., May 14, 2007; 35(9): 3076 - 3086. [Abstract] [Full Text] [PDF]

				S. A. N. McElhinny, C. M. Stith, P. M. J. Burgers, and T. A. Kunkel Inefficient Proofreading and Biased Error Rates during Inaccurate DNA Synthesis by a Mutant Derivative of Saccharomyces cerevisiae DNA Polymerase {delta} J. Biol. Chem., January 26, 2007; 282(4): 2324 - 2332. [Abstract] [Full Text] [PDF]