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J Biol Chem, Vol. 275, Issue 16, 11809-11813, April 21, 2000

Single Nucleotide Patch Base Excision Repair Is the Major Pathway for Removal of Thymine Glycol from DNA in Human Cell Extracts*

Grigory L. DianovDagger §, Tanja ThyboDagger ||, Irina I. DianovaDagger , Leonora J. LipinskiDagger , and Vilhelm A. BohrDagger

From the Dagger  Laboratory of Molecular Genetics, NIA, National Institutes of Health, Baltimore, Maryland 21224 and the  Department of Molecular and Structural Biology, Aarhus University, Denmark

The repair pathways involved in the removal of thymine glycol (TG) from DNA by human cell extracts have been examined. Closed circular DNA constructs containing a single TG at a defined site were used as substrates to determine the patch size generated after in vitro repair by cell extracts. Restriction analysis of the repair incorporation in the vicinity of the lesion indicated that the majority of TG was repaired through the base excision repair (BER) pathways. Repair incorporation 5' to the lesion, characteristic for the nucleotide excision repair pathway, was not found. More than 80% of the TG repair was accomplished by the single-nucleotide repair mechanism, and the remaining TGs were removed by the long patch BER pathway. We also analyzed the role of the xeroderma pigmentosum, complementation group G (XPG) protein in the excision step of BER. Cell extracts deficient in XPG protein had an average 25% reduction in TG incision. These data show that BER is the primary pathway for repair of TG in DNA and that XPG protein may be involved in repair of TG as an accessory factor.

* 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.

§ To whom correspondence should be addressed: Lab. of Molecular Genetics, NIA, NIH, 5600 Nathan Shock Dr., Baltimore, MD 21224. Tel.: 410-558-8562; Fax: 410-558-8157; E-mail: dianovg@grc.nia.nih. gov.

|| Supported by the Danish Center for Molecular Gerontology.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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