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

J. Biol. Chem., Vol. 283, Issue 25, 17075-17082, June 20, 2008

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 283/25/17075    most recent M801238200v1 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 Minko, I. G. Articles by Lloyd, R. S. Search for Related Content PubMed PubMed Citation Articles by Minko, I. G. Articles by Lloyd, R. S. Social Bookmarking                      What's this?

Role for DNA Polymerase in the Processing of N²-N²-Guanine Interstrand Cross-links^*

Irina G. Minko¹, Michael B. Harbut¹², Ivan D. Kozekov, Albena Kozekova, Petra M. Jakobs^¶, Susan B. Olson^¶, Robb E. Moses^¶, Thomas M. Harris, Carmelo J. Rizzo, and R. Stephen Lloyd^¶3

From the Center for Research on Occupational and Environmental Toxicology and ^¶Department of Molecular and Medical Genetics, Oregon Health & Science University, Portland, Oregon 97239 and the Department of Chemistry, Center in Molecular Toxicology, Vanderbilt University, Nashville, Tennessee 37235

Although there exists compelling genetic evidence for a homologous recombination-independent pathway for repair of interstrand cross-links (ICLs) involving translesion synthesis (TLS), biochemical support for this model is lacking. To identify DNA polymerases that may function in TLS past ICLs, oligodeoxynucleotides were synthesized containing site-specific ICLs in which the linkage was between N²-guanines, similar to cross-links formed by mitomycin C and enals. Here, data are presented that mammalian cell replication of DNAs containing these lesions was 97% accurate. Using a series of oligodeoxynucleotides that mimic potential intermediates in ICL repair, we demonstrate that human polymerase (pol) not only catalyzed accurate incorporation opposite the cross-linked guanine but also replicated beyond the lesion, thus providing the first biochemical evidence for TLS past an ICL. The efficiency of TLS was greatly enhanced by truncation of both the 5 ' and 3 ' ends of the nontemplating strand. Further analyses showed that although yeast Rev1 could incorporate a dCTP opposite the cross-linked guanine, no evidence was found for TLS by pol or a pol /Rev1 combination. Because pol was able to bypass these ICLs, biological evidence for a role for pol in tolerating the N²-N²-guanine ICLs was sought; both cell survival and chromosomal stability were adversely affected in pol -depleted cells following mitomycin C exposure. Thus, biochemical data and cellular studies both suggest a role for pol in the processing of N²-N²-guanine ICLs.

Received for publication, February 14, 2008 , and in revised form, April 14, 2008.

^* This work was supported, in whole or in part, by National Institutes of Health Grant ES 05355 and Center Grant ES 000267. 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.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Methods, additional references, Table 1, and Fig. 1.

¹ Both authors contributed equally to this work.

² Present address: Graduate Group in Pharmacological Sciences, University of Pennsylvania, Philadelphia, PA 19104.

³ To whom correspondence should be addressed: Center for Research on Occupational and Environmental Toxicology, Oregon Health and Science University, L606, 3181 SW Sam Jackson Park Rd., Portland, OR 97239. Tel.: 503-494-9957; Fax: 503-494-6831; E-mail: lloydst{at}ohsu.edu.

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

This article has been cited by other articles:

				S. Couve, G. Mace-Aime, F. Rosselli, and M. K. Saparbaev The Human Oxidative DNA Glycosylase NEIL1 Excises Psoralen-induced Interstrand DNA Cross-links in a Three-stranded DNA Structure J. Biol. Chem., May 1, 2009; 284(18): 11963 - 11970. [Abstract] [Full Text] [PDF]

				H. Zhang, R. L. Eoff, I. D. Kozekov, C. J. Rizzo, M. Egli, and F. P. Guengerich Versatility of Y-family Sulfolobus solfataricus DNA Polymerase Dpo4 in Translesion Synthesis Past Bulky N2-Alkylguanine Adducts J. Biol. Chem., February 6, 2009; 284(6): 3563 - 3576. [Abstract] [Full Text] [PDF]

				A. Kumari, I. G. Minko, M. B. Harbut, S. E. Finkel, M. F. Goodman, and R. S. Lloyd Replication Bypass of Interstrand Cross-link Intermediates by Escherichia coli DNA Polymerase IV J. Biol. Chem., October 10, 2008; 283(41): 27433 - 27437. [Abstract] [Full Text] [PDF]