Article
- The EMBO Journal (2009) 28, 621 - 631
- doi:10.1038/emboj.2009.27
Published online: 12 February 2009
There is a Have you seen ...? (March 2009) associated with this Article.
Subject Category:
K63-specific deubiquitination by two JAMM/MPN+ complexes: BRISC-associated Brcc36 and proteasomal Poh1
Eric M Cooper1, Colleen Cutcliffe1,a, Troels Z Kristiansen2, Akhilesh Pandey2, Cecile M Pickart1,b and Robert E Cohen1
- Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA
- McKusick-Nathans Institute of Genetic Medicine and Department of Biological Chemistry, Johns Hopkins University, Baltimore, MD, USA
Correspondence to:
Eric M Cooper, Department of Biochemistry and Molecular Biology, Johns Hopkins University, Bloomberg School of Public Health, 615 N. Wolfe St, Baltimore, MD 21205, USA. Tel.: +410 955 2926; Fax: +410 614 3734; E-mail: emcooper@jhsph.edu
Robert E Cohen, Department of Biochemistry and Molecular Biology, Johns Hopkins University, Bloomberg School of Public Health, 615 N. Wolfe St, Baltimore, MD 21205, USA. Tel.: +410 955 2926; Fax: +410 614 3734; E-mail: rcohen@jhsph.edu
aPresent address: Elan Pharmaceuticals, South San Francisco, CA 94080, USA
bDeceased
Received 18 June 2008; Accepted 19 January 2009
Abstract
An unusual deubiquitinating (DUB) activity exists in HeLa cell extracts that is highly specific for cleaving K63-linked but not K48-linked polyubiquitin chains. The activity is insensitive to both N-ethyl-maleimide and ubiquitin aldehyde, indicating that it lacks an active site cysteine residue, and gel filtration experiments show that it resides in a high molecular weight (
600 kDa) complex. Using a biochemical approach, we found that the K63-specific DUB activity co-fractionated through seven chromatographic steps with three multisubunit complexes: the 19S (PA700) portion of the 26S proteasome, the COP9 signalosome (CSN) and a novel complex that includes the JAMM/MPN+ domain-containing protein Brcc36. When we analysed the individual complexes, we found that the activity was intrinsic to PA700 and the Brcc36 isopeptidase complex (BRISC), but that the CSN-associated activity was due entirely to an interaction with Brcc36. None of the complexes cleave K6, K11, K29, K48 or 
-linked polyubiquitin, but they do cleave K63 linkages within mixed-linkage chains. Our results suggest that specificity for K63-linked polyubiquitin is a common property of the JAMM/MPN+ family of DUBs.
Keywords:
- Brcc36,
- deubiquitinating enzyme,
- isopeptidase,
- JAMM,
- Poh1
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