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

J. Biol. Chem., Vol. 278, Issue 21, 19079-19086, May 23, 2003

This Article Full Text Full Text (PDF) All Versions of this Article: 278/21/19079    most recent M208604200v1 Alert me when this article is cited 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 Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Limsirichaikul, S. Articles by Suzuki, M. Search for Related Content PubMed PubMed Citation Articles by Limsirichaikul, S. Articles by Suzuki, M. Social Bookmarking                      What's this?

The Gly-952 Residue of Saccharomyces cerevisiae DNA Polymerase Is Important in Discriminating Correct Deoxyribonucleotides from Incorrect Ones^*

Siripan Limsirichaikul , Masanori Ogawa , Atsuko Niimi , Shigenori Iwai , Takashi Murate ¶, Shonen Yoshida  and Motoshi Suzuki  ||

From the Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, ^¶ Nagoya University School of Health Science, Nagoya 461-8673, Japan

Gly-952 is a conserved residue in Saccharomyces cerevisiae DNA polymerase (pol ) that is strictly required for catalytic activity and for genetic complementation of a pol -deficient yeast strain. This study analyzes the role of Gly-952 by characterizing the biochemical properties of Gly-952 mutants. Analysis of the nucleotide incorporation specificity of pol G952A showed that this mutant incorporates nucleotides with extraordinarily low fidelity. In a steady-state kinetic assay to measure nucleotide misincorporation, pol G952A incorporated incorrect nucleotides more efficiently than correct nucleotides opposite template C, G, and T. The fidelity of the G952A mutant polymerase was highest at template A, where the ratio of incorporation of dCMP to dTMP was as high as 0.37. Correct nucleotide insertion was 500- to 3500-fold lower for G952A than for wild type pol , with up to 22-fold increase in pyrimidine misincorporation. The K_m for G952A pol bound to mismatched termini T:T, T:C, C:A, and A:C was 71- to 460-fold lower than to a matched terminus. Furthermore, pol G952A preferentially incorporated pyrimidine instead of dAMP opposite an abasic site, cis-syn cyclobutane di-thymine, or (6–4) di-thymine photoproduct. These data demonstrate that Gly-952 is a critical residue for catalytic efficiency and error prevention in S. cerevisiae pol .

Received for publication, August 22, 2002 , and in revised form, February 5, 2003.

^|| To whom correspondence should be addressed. Tel.: 81-52-744-2456; Fax: 81-52-744-2457; E-mail: msuzuki{at}med.nagoya-u.ac.jp.

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

This article has been cited by other articles:

				J. M. Barrero, R. Gonzalez-Bayon, J. C. del Pozo, M. R. Ponce, and J. L. Micol INCURVATA2 Encodes the Catalytic Subunit of DNA Polymerase {alpha} and Interacts with Genes Involved in Chromatin-Mediated Cellular Memory in Arabidopsis thaliana PLANT CELL, September 1, 2007; 19(9): 2822 - 2838. [Abstract] [Full Text] [PDF]

				A. Niimi, S. Limsirichaikul, S. Yoshida, S. Iwai, C. Masutani, F. Hanaoka, E. T. Kool, Y. Nishiyama, and M. Suzuki Palm Mutants in DNA Polymerases {alpha} and {eta} Alter DNA Replication Fidelity and Translesion Activity Mol. Cell. Biol., April 1, 2004; 24(7): 2734 - 2746. [Abstract] [Full Text] [PDF]

				M. Ogawa, S. Limsirichaikul, A. Niimi, S. Iwai, S. Yoshida, and M. Suzuki Distinct Function of Conserved Amino Acids in the Fingers of Saccharomyces cerevisiae DNA Polymerase {alpha} J. Biol. Chem., May 23, 2003; 278(21): 19071 - 19078. [Abstract] [Full Text] [PDF]