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Originally published In Press as doi:10.1074/jbc.M709275200 on December 26, 2007
J. Biol. Chem., Vol. 283, Issue 8, 4658-4664, February 22, 2008
Requirements for the Interaction of Mouse Pol with Ubiquitin and Its Biological Significance*
Caixia Guo ,
Tie-Shan Tang ,
Marzena Bienko¶1,
Ivan Dikic¶, and
Errol C. Friedberg 2
From the
Laboratory of Molecular Pathology, Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9072, the Department of Physiology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, and the ¶Institute for Biochemistry II, Goethe University Medical School, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany
Pol protein is a eukaryotic member of the DinB/Pol branch of the Y-family DNA polymerases, which are involved in the tolerance of DNA damage by replicative bypass. Despite universal conservation through evolution, the precise role(s) of Pol in this process has remained unknown. Here we report that mouse Pol can physically interact with ubiquitin by yeast two-hybrid screening, glutathione S-transferase pulldown, and immunoprecipitation methods. The association of Pol with ubiquitin requires the ubiquitin-binding motifs located at the C terminus of Pol . In addition, Pol binds with monoubiquitinated proliferating cell nuclear antigen (PCNA) more robustly than with non-ubiquitinated PCNA. The ubiquitin-binding motifs mediate the enhanced association between monoubiquitinated PCNA and Pol . The ubiquitin-binding motifs are also required for Pol to form nuclear foci after UV radiation. However, the ubiquitin-binding motifs do not affect Pol half-life. Finally, we have examined levels of Pol expression following the exposure of mouse cells to benzo[a]pyrene-dihydrodiol epoxide or UVB radiation.
Received for publication, November 12, 2007
, and in revised form, December 20, 2007.
* This study was supported in part by Grants ES11344 from NIEHS, National Institutes of Health (to E. C. F.), and DI 931/1-1 from the Deutsche Forschungsgemeinschaft (to I. D.). 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.
1 Supported by the Ernst Schering Foundation.
2 To whom correspondence should be addressed. Tel.: 214-648-4020; Fax: 214-648-4067; E-mail: friedberg.errol{at}pathology.swmed.edu.

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Copyright © 2008 by the American Society for Biochemistry and Molecular Biology.
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