title: Structural and kinetic analysis of the COP9-Signalosome activation and the cullin-RING ubiquitin ligase deneddylation cycle.
abstract: The COP9-Signalosome (CSN) regulates cullin-RING ubiquitin ligase (CRL) activity and assembly by cleaving Nedd8 from cullins. Free CSN is autoinhibited, and it remains unclear how it becomes activated. We combine structural and kinetic analyses to identify mechanisms that contribute to CSN activation and Nedd8 deconjugation. Both CSN and neddylated substrate undergo large conformational changes upon binding, with important roles played by the N-terminal domains of Csn2 and Csn4 and the RING domain of Rbx1 in enabling formation of a high affinity, fully active complex. The RING domain is crucial for deneddylation, and works in part through conformational changes involving insert-2 of Csn6. Nedd8 deconjugation and re-engagement of the active site zinc by the autoinhibitory Csn5 glutamate-104 diminish affinity for Cul1/Rbx1 by ~100-fold, resulting in its rapid ejection from the active site. Together, these mechanisms enable a dynamic deneddylation-disassembly cycle that promotes rapid remodeling of the cellular CRL network.
authors.affiliation: Keck School of Medicine, University of Southern California, Los Angeles, United States.|Combined MD/PhD Program, California Institute of Technology, Pasadena, United States.|Division of Biology and Biological Engineering, California Instittute of Technology, Pasadena, United States.
authors.forename: Ruzbeh
authors.initials: R
authors.lastname: Mosadeghi
authors.affiliation: Division of Biology and Biological Engineering, California Instittute of Technology, Pasadena, United States.
authors.forename: Kurt M
authors.initials: KM
authors.lastname: Reichermeier
authors.affiliation: Department of Biology, Institute of Biochemistry, Swiss Federal Institute of Technology, Zurich, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Winkler
authors.affiliation: Department of Biology, Institute of Biochemistry, Swiss Federal Institute of Technology, Zurich, Switzerland.
authors.forename: Anne
authors.initials: A
authors.lastname: Schreiber
authors.affiliation: Division of Biology and Biological Engineering, California Instittute of Technology, Pasadena, United States.
authors.forename: Justin M
authors.initials: JM
authors.lastname: Reitsma
authors.affiliation: Division of Biology and Biological Engineering, California Instittute of Technology, Pasadena, United States.
authors.forename: Yaru
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Biology, Institute of Molecular Systems Biology, Swiss Federal Institute of Technology, Zürich, Switzerland.
authors.forename: Florian
authors.initials: F
authors.lastname: Stengel
authors.affiliation: Division of Biology and Biological Engineering, California Instittute of Technology, Pasadena, United States.
authors.forename: Junyue
authors.initials: J
authors.lastname: Cao
authors.affiliation: Division of Biology and Biological Engineering, California Instittute of Technology, Pasadena, United States.
authors.forename: Minsoo
authors.initials: M
authors.lastname: Kim
authors.affiliation: Proteome Exploration Lab, Beckman Institute, California Institute of Technology, Pasadena, United States.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Sweredoski
authors.affiliation: Proteome Exploration Lab, Beckman Institute, California Institute of Technology, Pasadena, United States.
authors.forename: Sonja
authors.initials: S
authors.lastname: Hess
authors.affiliation: Department of Biology, Institute of Molecular Systems Biology, Swiss Federal Institute of Technology, Zürich, Switzerland.
authors.forename: Alexander
authors.initials: A
authors.lastname: Leitner
authors.affiliation: Department of Biology, Institute of Molecular Systems Biology, Swiss Federal Institute of Technology, Zürich, Switzerland.|Faculty of Science, University of Zurich, Zurich, Switzerland.
authors.forename: Ruedi
authors.initials: R
authors.lastname: Aebersold
authors.affiliation: Department of Biology, Institute of Biochemistry, Swiss Federal Institute of Technology, Zurich, Switzerland.
authors.forename: Matthias
authors.identifier: 0000-0002-2160-6824
authors.initials: M
authors.lastname: Peter
authors.affiliation: Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States.|Howard Hughes Medical Institute, California Institute of Technology, Pasadena, United States.
authors.forename: Raymond J
authors.identifier: 0000-0002-3671-9354
authors.initials: RJ
authors.lastname: Deshaies
authors.affiliation: Department of Biology, Institute of Biochemistry, Swiss Federal Institute of Technology, Zurich, Switzerland.
authors.forename: Radoslav I
authors.initials: RI
authors.lastname: Enchev
mesh_terms: D020285:Cryoelectron Microscopy; D044842:Cullin Proteins / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D013058:Mass Spectrometry; D008958:Models, Molecular; D009097:Multienzyme Complexes / Q000737:chemistry / Q000378:metabolism*; D000075687:NEDD8 Protein; D010447:Peptide Hydrolases / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D025801:Ubiquitin / Q000378:metabolism*; D014452:Ubiquitins / Q000378:metabolism*
chemical_list: D044842:Cullin Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009097:Multienzyme Complexes; D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; D025801:Ubiquitin; D014452:Ubiquitins; D010447:Peptide Hydrolases
keywords: COP9; CRL; CSN; Nedd8; biochemistry; biophysics; cell biology; cullin; human; structural biology; ubiquitin