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

This Article Full Text Full Text (PDF) 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 Chagovetz, A. M. Articles by Preston, B. D. Search for Related Content PubMed PubMed Citation Articles by Chagovetz, A. M. Articles by Preston, B. D. Social Bookmarking                      What's this?

Volume 272, Number 44, Issue of October 31, 1997 pp. 27501-27504
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

COMMUNICATION:
Increased Activity and Fidelity of DNA Polymerase  on Single-nucleotide Gapped DNA

(Received for publication, July 31, 1997)

Alexander M. Chagovetz , Joann B. Sweasy ^§ and Bradley D. Preston

From the  Departments of Biochemistry and Radiation Oncology, Eccles Institute of Human Genetics and Huntsman Cancer Institute, University of Utah, Salt Lake City, Utah 84112 and the ^§ Departments of Therapeutic Radiology and Genetics, Yale University School of Medicine, New Haven, Connecticut 06520

DNA polymerase  (pol ) is an error-prone polymerase that plays a central role in mammalian base excision repair. To better characterize the mechanisms governing rat pol  activity, we examined polymerization on synthetic primer-templates of different structure. Steady-state kinetic analyses revealed that the catalytic efficiency of pol  (k_cat/K_m,dNTP^app) is strongly influenced by gap size and the presence of a phosphate group at the 5-margin of the gap. pol  exhibited the highest catalytic efficiency on 5-phosphorylated 1-nucleotide gapped DNA. This efficiency was 500 times higher than on non-phosphorylated 1-nucleotide and 6-nucleotide (with or without PO₄) gapped DNAs and 2,500 times higher than on primer-template with no gaps. The nucleotide insertion fidelity of pol , as judged by its ability to form G-N mispairs, was also higher (10-100 times) on 5-phosphorylated single-nucleotide gapped DNA compared with the other DNA substrates studied. These data suggest that a primary function of mammalian pol  is to fill 5-phosphorylated 1-nucleotide gaps.

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

This article has been cited by other articles:

				S. Dalal, A. Chikova, J. Jaeger, and J. B. Sweasy The Leu22Pro tumor-associated variant of DNA polymerase beta is dRP lyase deficient Nucleic Acids Res., February 2, 2008; 36(2): 411 - 422. [Abstract] [Full Text] [PDF]

				K. Chan, S. Houlbrook, Q.-M. Zhang, M. Harrison, I. D. Hickson, and G. L. Dianov Overexpression of DNA polymerase {beta} results in an increased rate of frameshift mutations during base excision repair Mutagenesis, May 1, 2007; 22(3): 183 - 188. [Abstract] [Full Text] [PDF]

				W. W. Duym, K. A. Fiala, N. Bhatt, and Z. Suo Kinetic Effect of a Downstream Strand and Its 5'-Terminal Moieties on Single Nucleotide Gap-filling Synthesis Catalyzed by Human DNA Polymerase {lambda} J. Biol. Chem., November 24, 2006; 281(47): 35649 - 35655. [Abstract] [Full Text] [PDF]

				B. A. Sokhansanj and D. M. Wilson III Estimating the effect of human base excision repair protein variants on the repair of oxidative DNA base damage. Cancer Epidemiol. Biomarkers Prev., May 1, 2006; 15(5): 1000 - 1008. [Abstract] [Full Text] [PDF]

				M. Redrejo-Rodriguez, R. Garcia-Escudero, R. J. Yanez-Munoz, M. L. Salas, and J. Salas African Swine Fever Virus Protein pE296R Is a DNA Repair Apurinic/Apyrimidinic Endonuclease Required for Virus Growth in Swine Macrophages. J. Virol., May 1, 2006; 80(10): 4847 - 4857. [Abstract] [Full Text] [PDF]

				K. K.L. Chan, Q.-M. Zhang, and G. L. Dianov Base excision repair fidelity in normal and cancer cells Mutagenesis, May 1, 2006; 21(3): 173 - 178. [Abstract] [Full Text] [PDF]

				C. J. Dobson and S. L. Allinson The phosphatase activity of mammalian polynucleotide kinase takes precedence over its kinase activity in repair of single strand breaks. Nucleic Acids Res., January 1, 2006; 34(8): 2230 - 2237. [Abstract] [Full Text] [PDF]

				J. F. Ruiz, D. Lucas, E. Garcia-Palomero, A. I. Saez, M. A. Gonzalez, M. A. Piris, A. Bernad, and L. Blanco Overexpression of human DNA polymerase {micro} (Pol {micro}) in a Burkitt's lymphoma cell line affects the somatic hypermutation rate Nucleic Acids Res., November 1, 2004; 32(19): 5861 - 5873. [Abstract] [Full Text] [PDF]

				D. Wong and B. Demple Modulation of the 5'-Deoxyribose-5-phosphate Lyase and DNA Synthesis Activities of Mammalian DNA Polymerase {beta} by Apurinic/Apyrimidinic Endonuclease 1 J. Biol. Chem., June 11, 2004; 279(24): 25268 - 25275. [Abstract] [Full Text] [PDF]

				A. Rasouli-Nia, F. Karimi-Busheri, and M. Weinfeld Stable down-regulation of human polynucleotide kinase enhances spontaneous mutation frequency and sensitizes cells to genotoxic agents PNAS, May 4, 2004; 101(18): 6905 - 6910. [Abstract] [Full Text] [PDF]

				S. Dalal, J. L. Kosa, and J. B. Sweasy The D246V Mutant of DNA Polymerase {beta} Misincorporates Nucleotides: EVIDENCE FOR A ROLE FOR THE FLEXIBLE LOOP IN DNA POSITIONING WITHIN THE ACTIVE SITE J. Biol. Chem., January 2, 2004; 279(1): 577 - 584. [Abstract] [Full Text] [PDF]

				D. Wong, M. S. DeMott, and B. Demple Modulation of the 3'->5'-Exonuclease Activity of Human Apurinic Endonuclease (Ape1) by Its 5'-incised Abasic DNA Product J. Biol. Chem., September 19, 2003; 278(38): 36242 - 36249. [Abstract] [Full Text] [PDF]

				M. Maitra, A. Gudzelak Jr., S.-X. Li, Y. Matsumoto, K. A. Eckert, J. Jager, and J. B. Sweasy Threonine 79 Is a Hinge Residue That Governs the Fidelity of DNA Polymerase beta by Helping to Position the DNA within the Active Site J. Biol. Chem., September 13, 2002; 277(38): 35550 - 35560. [Abstract] [Full Text] [PDF]

				H. Budworth, I. I. Dianova, V. N. Podust, and G. L. Dianov Repair of Clustered DNA Lesions. SEQUENCE-SPECIFIC INHIBITION OF LONG-PATCH BASE EXCISION REPAIR BY 8-OXOGUANINE J. Biol. Chem., June 7, 2002; 277(24): 21300 - 21305. [Abstract] [Full Text] [PDF]

				A. Skandalis and L. A. Loeb Enzymatic properties of rat DNA polymerase {beta} mutants obtained by randomized mutagenesis Nucleic Acids Res., June 1, 2001; 29(11): 2418 - 2426. [Abstract] [Full Text] [PDF]

				A. Tissier, J. P. McDonald, E. G. Frank, and R. Woodgate poliota , a remarkably error-prone human DNA polymerase Genes & Dev., July 1, 2000; 14(13): 1642 - 1650. [Abstract] [Full Text]

				A. Vaisman and S. G. Chaney The Efficiency and Fidelity of Translesion Synthesis past Cisplatin and Oxaliplatin GpG Adducts by Human DNA Polymerase beta J. Biol. Chem., April 21, 2000; 275(17): 13017 - 13025. [Abstract] [Full Text] [PDF]

				G. J. Klarmann, R. A. Smith, R. F. Schinazi, T. W. North, and B. D. Preston Site-specific Incorporation of Nucleoside Analogs by HIV-1 Reverse Transcriptase and the Template Grip Mutant P157S. TEMPLATE INTERACTIONS INFLUENCE SUBSTRATE RECOGNITION AT THE POLYMERASE ACTIVE SITE J. Biol. Chem., January 7, 2000; 275(1): 359 - 366. [Abstract] [Full Text] [PDF]

				J. L. Kosa and J. B. Sweasy The E249K Mutator Mutant of DNA Polymerase beta Extends Mispaired Termini J. Biol. Chem., December 10, 1999; 274(50): 35866 - 35872. [Abstract] [Full Text] [PDF]

				C. A. Clairmont, L. Narayanan, K.-W. Sun, P. M. Glazer, and J. B. Sweasy The Tyr-265-to-Cys mutator mutant of DNA polymerase beta induces a mutator phenotype in mouse LN12 cells PNAS, August 17, 1999; 96(17): 9580 - 9585. [Abstract] [Full Text] [PDF]

				Y. CANITROT, M. FRÉCHET, L. SERVANT, C. CAZAUX, and J.-S. HOFFMANN Overexpression of DNA polymerase ß: a genomic instability enhancer process FASEB J, June 1, 1999; 13(9): 1107 - 1111. [Abstract] [Full Text]

				E. Efrati, G. Tocco, R. Eritja, S. H. Wilson, and M. F. Goodman "Action-at-a-Distance" Mutagenesis. 8-OXO-7,8-DIHYDRO-2'-DEOXYGUANOSINE CAUSES BASE SUBSTITUTION ERRORS AT NEIGHBORING TEMPLATE SITES WHEN COPIED BY DNA POLYMERASE beta J. Biol. Chem., May 28, 1999; 274(22): 15920 - 15926. [Abstract] [Full Text] [PDF]

				W. P. Osheroff, H. K. Jung, W. A. Beard, S. H. Wilson, and T. A. Kunkel The Fidelity of DNA Polymerase beta  during Distributive and Processive DNA Synthesis J. Biol. Chem., February 5, 1999; 274(6): 3642 - 3650. [Abstract] [Full Text] [PDF]

				Z. Suo and K. A. Johnson Selective Inhibition of HIV-1 Reverse Transcriptase by an Antiviral Inhibitor, (R)-9-(2-Phosphonylmethoxypropyl)adenine J. Biol. Chem., October 16, 1998; 273(42): 27250 - 27258. [Abstract] [Full Text] [PDF]

				R. J. Sanderson and D. W. Mosbaugh Fidelity and Mutational Specificity of Uracil-initiated Base Excision DNA Repair Synthesis in Human Glioblastoma Cell Extracts J. Biol. Chem., September 18, 1998; 273(38): 24822 - 24831. [Abstract] [Full Text] [PDF]

				C. A. Clairmont and J. B. Sweasy The Pol beta -14 Dominant Negative Rat DNA Polymerase beta  Mutator Mutant Commits Errors during the Gap-Filling Step of Base Excision Repair in Saccharomyces cerevisiae J. Bacteriol., May 1, 1998; 180(9): 2292 - 2297. [Abstract] [Full Text]

				B. J. Vande Berg, W. A. Beard, and S. H. Wilson DNA Structure and Aspartate 276 Influence Nucleotide Binding to Human DNA Polymerase beta . IMPLICATION FOR THE IDENTITY OF THE RATE-LIMITING CONFORMATIONAL CHANGE J. Biol. Chem., January 26, 2001; 276(5): 3408 - 3416. [Abstract] [Full Text] [PDF]

				A. Vaisman, M. W. Warren, and S. G. Chaney The Effect of DNA Structure on the Catalytic Efficiency and Fidelity of Human DNA Polymerase beta on Templates with Platinum-DNA Adducts J. Biol. Chem., May 25, 2001; 276(22): 18999 - 19005. [Abstract] [Full Text] [PDF]

				A. M. Shah, S.-X. Li, K. S. Anderson, and J. B. Sweasy Y265H Mutator Mutant of DNA Polymerase beta . PROPER GEOMETRIC ALIGNMENT IS CRITICAL FOR FIDELITY J. Biol. Chem., March 30, 2001; 276(14): 10824 - 10831. [Abstract] [Full Text] [PDF]