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Originally published In Press as doi:10.1074/jbc.M206889200 on October 3, 2002

J. Biol. Chem., Vol. 277, Issue 50, 48434-48440, December 13, 2002
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Human DNA Polymerase lambda  Functionally and Physically Interacts with Proliferating Cell Nuclear Antigen in Normal and Translesion DNA Synthesis*

Giovanni MagaDagger §, Giuseppe Villani, Kristijan Ramadan||, Igor Shevelev||**, Nicolas Tanguy Le Gac, Luis BlancoDagger Dagger , Giuseppina BlancaDagger , Silvio SpadariDagger , and Ulrich Hübscher||

From the Dagger  Istituto di Genetica Molecolare-Consiglio Nazionale delle Ricerche, Via Abbiategrasso 207, I-27100 Pavia, Italy, the  Institut de Pharmacology et de Biologie Structurale, Centre National de la Recherche Scientifique, 205 Route de Narbonne, 31077 Toulouse Cedex, France, the || Institute of Veterinary Biochemistry and Molecular Biology, University of Zürich-Irchel, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland, the ** Department of Molecular and Radiation Biophysics, Petersburg Nuclear Physics Institute, Leningrad District, Gatchina 188300, Russia, and the Dagger Dagger  Centro di Biologia Molecular Severo Ochoa, Consejo Superior de Investigaciones Cientificas-Universitad Autonoma, Madrid 29089, Spain

Proliferating cell nuclear antigen (PCNA) has been shown to interact with a variety of DNA polymerases (pol) such as pol delta , pol epsilon , pol iota , pol kappa , pol eta , and pol beta . Here we show that PCNA directly interacts with the newly discovered pol lambda  cloned from human cells. This interaction stabilizes the binding of pol lambda  to the primer template, thus increasing its affinity for the hydroxyl primer and its processivity in DNA synthesis. However, no effect of PCNA was detected on the rate of nucleotide incorporation or discrimination efficiency by pol lambda . PCNA was found to stimulate efficient synthesis by pol lambda  across an abasic (AP) site. When compared with pol delta , human pol lambda  showed the ability to incorporate a nucleotide in front of the lesion. Addition of PCNA led to efficient elongation past the AP site by pol lambda  but not by pol delta . However, when tested on a template containing a bulky DNA lesion, such as the major cisplatin Pt-d(GpG) adduct, PCNA could not allow translesion synthesis by pol lambda . Our results suggest that the complex between PCNA and pol lambda  may play an important role in the bypass of abasic sites in human cells.

* This work was supported by the Consiglio Nazionale delle Ricerche Target Project on Biotechnology and CNR Agenzia 2000 (to S. S. and G. M.), by Swiss National Science Foundation Grant 31-61361.00 (to K. R.), by the Kanton of Zürich (to I. S., G. M. in part, and U. H.), and by Grant 4373 from the Association pour la Recherche sur le Cancer (to G. V.).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: IGM-CNR, Via Abbiategrasso 207, I-27100, Pavia, Italy. Tel.: 39-0382-546355; Fax: 39-0382-422286; E-mail: maga@igm.cnr.it.

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


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