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Originally published In Press as doi:10.1074/jbc.M207101200 on October 17, 2002

J. Biol. Chem., Vol. 277, Issue 51, 50046-50053, December 20, 2002
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A Role for DNA Polymerase beta  in Mutagenic UV Lesion Bypass*

Laurence ServantDagger §, Christophe CazauxDagger , Anne BiethDagger , Shigenori Iwai, Fumio Hanaoka||**, and Jean-Sébastien HoffmannDagger Dagger Dagger

From the Dagger  Group "Genetic instability and cancer" at the Institut de Pharmacologie et Biologie Structurale, UMR CNRS 5089, 31077 Toulouse cédex 4, France, the  Biomolecular Engineering Research Institute, 6-2-3 Furuedai, Suita, Osaka 565-0874, Japan, and the || Graduate School of Frontier Biosciences, Osaka University and Core Research for Engineering, Science, and Technology (CREST), Japan Science and Technology Corporation, 1-3 Yamada-oka, Suita, Osaka 565-0871, Japan

We report here that DNA polymerase beta  (pol beta ), the base excision repair polymerase, is highly expressed in human melanoma tissues, known to be associated with UV radiation exposure. To investigate the potential role of pol beta  in UV-induced genetic instability, we analyzed the cellular and molecular effects of excess pol beta . We firstly demonstrated that mammalian cells overexpressing pol beta  are resistant and hypermutagenic after UV irradiation and that replicative extracts from these cells are able to catalyze complete translesion replication of a thymine-thymine cyclobutane pyrimidine dimer (CPD). By using in vitro primer extension reactions with purified pol beta , we showed that CPD as well as, to a lesser extent, the thymine-thymine pyrimidine-pyrimidone (6-4) photoproduct, were bypassed. pol beta  mostly incorporates the correct dATP opposite the 3'-terminus of both CPD and the (6-4) photoproduct but can also misinsert dCTP at a frequency of 32 and 26%, respectively. In the case of CPD, efficient and error-prone extension of the correct dATP was found. These data support a biological role of pol beta  in UV lesion bypass and suggest that deregulated pol beta  may enhance UV-induced genetic instability.

* This work was exclusively supported financially by "La Ligue Nationale contre le Cancer" (Equipe labelisée).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.

§ An ARC fellowship recipient.

** To whom correspondence may be addressed. E-mail: fhanaoka@imcb.osaka-u.ac.jp.

Dagger Dagger To whom correspondence may be addressed. E-mail: jseb@ipbs.fr.

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


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