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Molecular and Cellular Biology, December 2006, p. 8892-8900, Vol. 26, No. 23
0270-7306/06/$08.00+0     doi:10.1128/MCB.01118-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Ubiquitin-Binding Motifs in REV1 Protein Are Required for Its Role in the Tolerance of DNA Damage{triangledown} ,{dagger}

Caixia Guo,1 Tie-Shan Tang,2 Marzena Bienko,3 Joanne L. Parker,4 Aleksandra B. Bielen,4 Eiichiro Sonoda,5 Shunichi Takeda,5 Helle D. Ulrich,4 Ivan Dikic,3 and Errol C. Friedberg1*

Laboratory of Molecular Pathology, Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9072,1 Department of Physiology, University of Texas Southwestern Medical Center, Dallas, Texas 75390,2 Institute for Biochemistry II, Goethe University Medical School, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany,3 Cancer Research United Kingdom, Clare Hall Laboratories, Blanche Lane, South Mimms EN6 3LD, United Kingdom,4 Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Kyoto 606-8501, Japan5

Received 21 June 2006/ Returned for modification 3 August 2006/ Accepted 6 September 2006

REV1 protein is a eukaryotic member of the Y family of DNA polymerases involved in the tolerance of DNA damage by replicative bypass. The precise role(s) of REV1 in this process is not known. Here we show, by using the yeast two-hybrid assay and the glutathione S-transferase pull-down assay, that mouse REV1 can physically interact with ubiquitin. The association of REV1 with ubiquitin requires the ubiquitin-binding motifs (UBMs) located at the C terminus of REV1. The UBMs also mediate the enhanced association between monoubiquitylated PCNA and REV1. In cells exposed to UV radiation, the association of REV1 with replication foci is dependent on functional UBMs. The UBMs of REV1 are shown to contribute to DNA damage tolerance and damage-induced mutagenesis in vivo.

* Corresponding author. Mailing address: Laboratory of Molecular Pathology, Department of Pathology, University of Texas Southwestern Medical Center, Dallas, TX 75390-9072. Phone: (214) 648-4020. Fax: (214) 648-4067. E-mail: friedberg.errol{at}pathology.swmed.edu.

{triangledown} Published ahead of print on 18 September 2006.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, December 2006, p. 8892-8900, Vol. 26, No. 23
0270-7306/06/$08.00+0     doi:10.1128/MCB.01118-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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