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Originally published In Press as doi:10.1074/jbc.M112297200 on January 30, 2002

J. Biol. Chem., Vol. 277, Issue 15, 12777-12783, April 12, 2002
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Transcription-coupled DNA Repair Is Genomic Context-dependent*

Zhaohui FengDagger §, Wenwei HuDagger §, Elena KomissarovaDagger , Annie Pao, Mien-Chie Hung, Gerald M. Adair||, and Moon-shong TangDagger **

From the Dagger  Department of Environmental Medicine, Pathology and Medicine, New York University School of Medicine, Tuxedo, New York 10987, the  Department of Molecular and Cellular Oncology, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, and the || Department of Carcinogenesis, University of Texas M. D. Anderson Cancer Center, Smithville, Texas 78957

DNA damage is preferentially repaired in the transcribed strand of many active genes. Although the concept of DNA repair coupled with transcription has been widely accepted, its mechanisms remain elusive. We recently reported that in Chinese hamster ovary cells while ultraviolet light-induced cyclobutane pyrimidine dimers (CPDs) are preferentially repaired in the transcribed strand of dihydrofolate reductase gene, CPDs are efficiently repaired in both strands of adenine phosphoribosyltransferase (APRT) locus, in either a transcribed or nontranscribed APRT gene (1). These results suggested that the transcription dependence of repair may depend on genomic context. To test this hypothesis, we constructed transfectant cell lines containing a single, actively transcribed APRT gene, integrated at different genomic sites. Mapping of CPD repair in the integrated APRT genes in three transfectant cell lines revealed two distinct repair patterns, either preferential repair of CPDs in the transcribed strand or very poor repair in both strands. Similar kinetics of micrococcal nuclease digestion were seen for all three transfectant APRT gene domains and endogenous APRT locus. Our results suggest that both the efficiency and strand-specificity of repair of an actively transcribed gene are profoundly affected by genomic context but do not reflect changes in first order nucleosomal structure.

* This work was supported by Public Health Service Grants ES03124, ES08389 (M. -s. Tang) and GM56165 (G. M. Adair) from the National Institutes of Health and service core support from National Institute of Environmental Health Sciences Grants ES077784 and ES00260.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Both authors contributed equally to this work.

** To whom correspondence should be addressed. Tel.: 845-731-3585; Fax: 845-351-2385; E-mail: tang@env.med.nyu.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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