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Nucleic Acids Research Advance Access published online on October 25, 2008
Nucleic Acids Research, doi:10.1093/nar/gkn651
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© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Genome integrity, repair and replication

REV1 restrains DNA polymerase {zeta} to ensure frame fidelity during translesion synthesis of UV photoproducts in vivo

Dávid Szüts1, Adam P. Marcus1, Masayuki Himoto2, Shigenori Iwai2 and Julian E. Sale1,*

1Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 0QH, UK and 2Division of Chemistry, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan

*To whom correspondence should be addressed. Tel: +44 0 1223 252941; Fax: +44 01223 412178; Email: jes{at}mrc-lmb.cam.ac.uk

Received August 14, 2008. Revised September 11, 2008. Accepted September 18, 2008.

Exposure to ultraviolet light induces a number of forms of damage in DNA, of which (6–4) photoproducts present the most formidable challenge to DNA replication. No single DNA polymerase has been shown to bypass these lesions efficiently in vitro suggesting that the coordinate use of a number of different enzymes is required in vivo. To further understand the mechanisms and control of lesion bypass in vivo, we have devised a plasmid-based system to study the replication of site-specific T–T(6–4) photoproducts in chicken DT40 cells. We show that DNA polymerase {zeta} is absolutely required for translesion synthesis (TLS) of this lesion, while loss of DNA polymerase {eta} has no detectable effect. We also show that either the polymerase-binding domain of REV1 or ubiquitinated PCNA is required for the recruitment of Pol{zeta} as the catalytic TLS polymerase. Finally, we demonstrate a previously unappreciated role for REV1 in ensuring bypass synthesis remains in frame with the template. Our data therefore suggest that REV1 not only helps to coordinate the delivery of DNA polymerase {zeta} to a stalled primer terminus but also restrains its activity to ensure that nucleotides are incorporated in register with the template strand.

Present address: Dávid Szüts, Division of Basic Medical Sciences, St George's, University of London, Cranmer Terrace, London SW17 0RE, UK


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