Higher-Order Assembly of BRCC36-KIAA0157 Is Required for DUB Activity and Biological Function.
BRCC36 is a Zn(2+)-dependent deubiquitinating enzyme (DUB) that hydrolyzes lysine-63-linked ubiquitin chains as part of distinct macromolecular complexes that participate in either interferon signaling or DNA-damage recognition. The MPN(+) domain protein BRCC36 associates with pseudo DUB MPN(-) proteins KIAA0157 or Abraxas, which are essential for BRCC36 enzymatic activity. To understand the basis for BRCC36 regulation, we have solved the structure of an active BRCC36-KIAA0157 heterodimer and an inactive BRCC36 homodimer. Structural and functional characterizations show how BRCC36 is switched to an active conformation by contacts with KIAA0157. Higher-order association of BRCC36 and KIAA0157 into a dimer of heterodimers (super dimers) was required for DUB activity and interaction with targeting proteins SHMT2 and RAP80. These data provide an explanation of how an inactive pseudo DUB allosterically activates a cognate DUB partner and implicates super dimerization as a new regulatory mechanism underlying BRCC36 DUB activity, subcellular localization, and biological function.
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 1090, Toronto, ON M5G 1X5, Canada.
Elton
E
Zeqiraj
Departments of Cancer Biology and Pathology, Abramson Family Cancer Research Institute, Basser Research Center for BRCA, Perelman School of Medicine, University of Pennsylvania, 421 Curie Boulevard, Philadelphia, PA 19104-6160, USA.
Lei
L
Tian
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 1090, Toronto, ON M5G 1X5, Canada.
Christopher A
CA
Piggott
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON L8S 4K1, Canada.
Monica C
MC
Pillon
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 1090, Toronto, ON M5G 1X5, Canada.
Nicole M
NM
Duffy
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 1090, Toronto, ON M5G 1X5, Canada.
Derek F
DF
Ceccarelli
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 1090, Toronto, ON M5G 1X5, Canada; Departments of Biochemistry and Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada.
Alexander F A
AF
Keszei
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 1090, Toronto, ON M5G 1X5, Canada.
Kristina
K
Lorenzen
Department of Chemistry and Chemical Biology, NE-CAT, Cornell University, Argonne, IL 60439, USA.
Igor
I
Kurinov
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 1090, Toronto, ON M5G 1X5, Canada.
Stephen
S
Orlicky
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 1090, Toronto, ON M5G 1X5, Canada.
Gerald D
GD
Gish
Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Science, Utrecht University, Padualaan 8, 3584 CH, Utrecht, the Netherlands.
Albert J R
AJ
Heck
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON L8S 4K1, Canada.
Alba
A
Guarné
Departments of Cancer Biology and Pathology, Abramson Family Cancer Research Institute, Basser Research Center for BRCA, Perelman School of Medicine, University of Pennsylvania, 421 Curie Boulevard, Philadelphia, PA 19104-6160, USA. Electronic address: rogergr@mail.med.upenn.edu.
Roger A
RA
Greenberg
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Avenue, Room 1090, Toronto, ON M5G 1X5, Canada; Departments of Biochemistry and Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada. Electronic address: sicheri@lunenfeld.ca.
Frank
F
Sicheri
D000818:Animals; D001000:Ants / Q000201:enzymology*; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D000072017:Deubiquitinating Enzymes; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D019476:Insect Proteins / Q000737:chemistry* / Q000502:physiology; D007700:Kinetics; D008565:Membrane Proteins / Q000737:chemistry; D008958:Models, Molecular; D034521:Nuclear Matrix-Associated Proteins / Q000737:chemistry* / Q000502:physiology; D011485:Protein Binding; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D064570:Ubiquitin-Specific Proteases / Q000737:chemistry* / Q000502:physiology
C577338:ABRAXAS2 protein, human; D019476:Insect Proteins; D008565:Membrane Proteins; D034521:Nuclear Matrix-Associated Proteins; C518017:BRCC3 protein, human; D000072017:Deubiquitinating Enzymes; D064570:Ubiquitin-Specific Proteases