This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Niimi, A.
Right arrow Articles by Suzuki, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Niimi, A.
Right arrow Articles by Suzuki, M.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, April 2004, p. 2734-2746, Vol. 24, No. 7
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.7.2734-2746.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Palm Mutants in DNA Polymerases {alpha} and {eta} Alter DNA Replication Fidelity and Translesion Activity

Atsuko Niimi,1 Siripan Limsirichaikul,1 Shonen Yoshida,1,{dagger} Shigenori Iwai,2 Chikahide Masutani,3 Fumio Hanaoka,3,4 Eric T. Kool,5 Yukihiro Nishiyama,6 and Motoshi Suzuki1*

Division of Molecular Carcinogenesis, Center for Neural Disease and Cancer,1 Department of Virology, Nagoya University Graduate School of Medicine, Nagoya 466-8550,6 Division of Chemistry, Graduate School of Engineering Science, Osaka University, Toyonaka, Osaka 560-8531,2 Graduate School of Frontier Biosciences, Osaka University, and Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, Suita, Osaka 565-0871,3 Cellular Physiology Laboratory, RIKEN, Wako, Saitama 351-0198, Japan,4 Department of Chemistry, Stanford University, Stanford, California 943055

Received 8 September 2003/ Returned for modification 18 October 2003/ Accepted 11 December 2003

We isolated active mutants in Saccharomyces cerevisiae DNA polymerase {alpha} that were associated with a defect in error discrimination. Among them, L868F DNA polymerase {alpha} has a spontaneous error frequency of 3 in 100 nucleotides and 570-fold lower replication fidelity than wild-type (WT) polymerase {alpha}. In vivo, mutant DNA polymerases confer a mutator phenotype and are synergistic with msh2 or msh6, suggesting that DNA polymerase {alpha}-dependent replication errors are recognized and repaired by mismatch repair. In vitro, L868F DNA polymerase {alpha} catalyzes efficient bypass of a cis-syn cyclobutane pyrimidine dimer, extending the 3' T 26,000-fold more efficiently than the WT. Phe34 is equivalent to residue Leu868 in translesion DNA polymerase {eta}, and the F34L mutant of S. cerevisiae DNA polymerase {eta} has reduced translesion DNA synthesis activity in vitro. These data suggest that high-fidelity DNA synthesis by DNA polymerase {alpha} is required for genomic stability in yeast. The data also suggest that the phenylalanine and leucine residues in translesion and replicative DNA polymerases, respectively, might have played a role in the functional evolution of these enzyme classes.

* Corresponding author. Mailing address: Division of Molecular Carcinogenesis, Center for Neural Disease and Cancer, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan. Phone: 81 52 744 2456. Fax: 81 52 744 2457. E-mail: msuzuki{at}med.nagoya-u.ac.jp.

{dagger} Present address: Kaikokai Rehabilitation Hospital, Amagun, Aichi 490-1405, Japan.

Molecular and Cellular Biology, April 2004, p. 2734-2746, Vol. 24, No. 7
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.7.2734-2746.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Stone, J. E., Kissling, G. E., Lujan, S. A., Rogozin, I. B., Stith, C. M., Burgers, P. M. J., Kunkel, T. A. (2009). Low-fidelity DNA synthesis by the L979F mutator derivative of Saccharomyces cerevisiae DNA polymerase {zeta}. Nucleic Acids Res 37: 3774-3787 [Abstract] [Full Text]  
  • Sabouri, N., Viberg, J., Goyal, D. K., Johansson, E., Chabes, A. (2008). Evidence for lesion bypass by yeast replicative DNA polymerases during DNA damage. Nucleic Acids Res 36: 5660-5667 [Abstract] [Full Text]  
  • Zhong, X., Pedersen, L. C., Kunkel, T. A. (2008). Characterization of a replicative DNA polymerase mutant with reduced fidelity and increased translesion synthesis capacity. Nucleic Acids Res 36: 3892-3904 [Abstract] [Full Text]  
  • Venkatesan, R. N., Treuting, P. M., Fuller, E. D., Goldsby, R. E., Norwood, T. H., Gooley, T. A., Ladiges, W. C., Preston, B. D., Loeb, L. A. (2007). Mutation at the Polymerase Active Site of Mouse DNA Polymerase {delta} Increases Genomic Instability and Accelerates Tumorigenesis. Mol. Cell. Biol. 27: 7669-7682 [Abstract] [Full Text]  
  • Barrero, J. M., Gonzalez-Bayon, R., del Pozo, J. C., Ponce, M. R., Micol, J. L. (2007). INCURVATA2 Encodes the Catalytic Subunit of DNA Polymerase {alpha} and Interacts with Genes Involved in Chromatin-Mediated Cellular Memory in Arabidopsis thaliana. Plant Cell 19: 2822-2838 [Abstract] [Full Text]  
  • Pursell, Z. F., Isoz, I., Lundstrom, E.-B., Johansson, E., Kunkel, T. A. (2007). 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 35: 3076-3086 [Abstract] [Full Text]  
  • McElhinny, S. A. N., Stith, C. M., Burgers, P. M. J., Kunkel, T. A. (2007). Inefficient Proofreading and Biased Error Rates during Inaccurate DNA Synthesis by a Mutant Derivative of Saccharomyces cerevisiae DNA Polymerase {delta}. J. Biol. Chem. 282: 2324-2332 [Abstract] [Full Text]  
  • Masuda, Y., Kamiya, K. (2006). Role of Single-stranded DNA in Targeting REV1 to Primer Termini. J. Biol. Chem. 281: 24314-24321 [Abstract] [Full Text]  
  • Venkatesan, R. N., Hsu, J. J., Lawrence, N. A., Preston, B. D., Loeb, L. A. (2006). Mutator Phenotypes Caused by Substitution at a Conserved Motif A Residue in Eukaryotic DNA Polymerase {delta}. J. Biol. Chem. 281: 4486-4494 [Abstract] [Full Text]  
  • Li, L., Murphy, K. M., Kanevets, U., Reha-Krantz, L. J. (2005). Sensitivity to Phosphonoacetic Acid: A New Phenotype to Probe DNA Polymerase {delta} in Saccharomyces cerevisiae. Genetics 170: 569-580 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»