Article
- The EMBO Journal (2009) 28, 2461 - 2468
- doi:10.1038/emboj.2009.160
Published online: 18 June 2009
There is a Have you seen ...? (August 2009) associated with this Article.
Subject Category:
Structural basis for specific recognition of Lys 63-linked polyubiquitin chains by tandem UIMs of RAP80
Yusuke Sato1,2, Azusa Yoshikawa1,2, Hisatoshi Mimura1, Masami Yamashita1,3, Atsushi Yamagata1 and Shuya Fukai1,3
- Structural Biology Laboratory, Life Science Division, Synchrotron Radiation Research Organization and Institute of Molecular and Cellular Biosciences, The University of Tokyo, Tokyo, Japan
- Department of Biological Information, Tokyo Institute of Technology, Yokohama, Japan
- Department of Medical Genome Sciences, Graduate School of Frontier Sciences, the University of Tokyo, Chiba, Japan
Correspondence to:
Shuya Fukai, Institute of Molecular and Cellular Biosciences, The University of Tokyo, General Research Bldg 211, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan. Tel.: +81 3 5841 7807; Fax: +81 3 5841 7807; E-mail: fukai@iam.u-tokyo.ac.jp
Received 20 April 2009; Accepted 20 May 2009
Abstract
RAP80 has a key role in the recruitment of the Abraxas–BRCC36–BRCA1–BARD1 complex to DNA-damage foci for DNA repair through specific recognition of Lys 63-linked polyubiquitinated proteins by its tandem ubiquitin-interacting motifs (UIMs). Here, we report the crystal structure of the RAP80 tandem UIMs (RAP80-UIM1-UIM2) in complex with Lys 63-linked di-ubiquitin at 2.2 Å resolution. The two UIMs, UIM1 and UIM2, and the 
-helical inter-UIM region together form a continuous 60 Å-long -helix. UIM1 and UIM2 bind to the proximal and distal ubiquitin moieties, respectively. Both UIM1 and UIM2 of RAP80 recognize an Ile 44-centered hydrophobic patch on ubiquitin but neither UIM interacts with the Lys 63-linked isopeptide bond. Our structure suggests that the inter-UIM region forms a 12 Å-long -helix that ensures that the UIMs are arranged to enable specific binding of Lys 63-linked di-ubiquitin. This was confirmed by pull-down analyses using RAP80-UIM1-UIM2 mutants of various length inter-UIM regions. Further, we show that the Epsin1 tandem UIM, which has an inter-UIM region similar to that of RAP80-UIM1-UIM2, also selectively binds Lys 63-linked di-ubiquitin.
Keywords:
- crystallography,
- DNA repair,
- ubiquitin,
- ubiquitin-interacting motif
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