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First published online 21 November 2006
doi: 10.1242/jcs.03291

Journal of Cell Science 119, 5137-5146 (2006)
Published by The Company of Biologists 2006
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Research Article

Werner syndrome protein participates in a complex with RAD51, RAD54, RAD54B and ATR in response to ICL-induced replication arrest

Marit Otterlei1,2,*, Per Bruheim1,3,§, Byungchan Ahn1,4,§, Wendy Bussen5, Parimal Karmakar1,6, Kathy Baynton7 and Vilhelm A. Bohr1

1 Laboratory of Molecular Gerontology, National Institute on Aging, NIH, 5600 Nathan Shock Dr., Baltimore, MD 21224, USA
2 Department of Cancer Research and Molecular Medicine, Laboratory Centre, Faculty of Medicine, Norwegian University of Science and Technology, Erling Skjalgsons gt. 1, N-7006 Trondheim, Norway
3 Department of Biotechnology, Norwegian University of Science and Technology, N-7491 Trondheim, Norway
4 Department of Life Sciences, University of Ulsan, Ulsan 680-749, Korea
5 Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, 333 Cedar St, SHM-C130, New Haven, CT 06515, USA
6 Department of Life Science and Biotechnology, Jadavpur University, Kolkata-700 032, WB, India
7 Research Center, Ste. Justine's Hospital, 3175 Cote Ste. Catherine, Montreal, Quebec H3T 1C5, Canada

* Author for correspondence (e-mail: marit.otterlei{at}ntnu.no)

Accepted 5 October 2006

Werner syndrome (WS) is a rare genetic disorder characterized by genomic instability caused by defects in the WRN gene encoding a member of the human RecQ helicase family. RecQ helicases are involved in several DNA metabolic pathways including homologous recombination (HR) processes during repair of stalled replication forks. Following introduction of interstrand DNA crosslinks (ICL), WRN relocated from nucleoli to arrested replication forks in the nucleoplasm where it interacted with the HR protein RAD52. In this study, we use fluorescence resonance energy transfer (FRET) and immune-precipitation experiments to demonstrate that WRN participates in a multiprotein complex including RAD51, RAD54, RAD54B and ATR in cells where replication has been arrested by ICL. We verify the WRN-RAD51 and WRN-RAD54B direct interaction in vitro. Our data support a role for WRN also in the recombination step of ICL repair.

Key words: WRN, ICL, RAD51, RAD54B, ATR, DNA repair




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© The Company of Biologists Ltd 2006