Structural and kinetic analysis of the COP9-Signalosome activation and the cullin-RING ubiquitin ligase deneddylation cycle

Mosadeghi, Ruzbeh; Reichermeier, Kurt M.; Winkler, Martin; Schreiber, Anne; Reitsma, Justin M.; Zhang, Yaru; Stengel, Florian; Cao, Junyue; Kim, Minsoo; Sweredoski, Michael J.; Hess, Sonja; Leitner, Alexander; Aebersold, Ruedi; Peter, Matthias; Deshaies, Raymond J.; Enchev, Radoslav I.

doi:10.7554/elife.12102

Cited by 82 publications

(71 citation statements)

References 48 publications

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“…In our EM maps and the other published CSN-CRL structures 16,19,20 , the exact position of NEDD8 and the Cullin-2 Winged-Helix B (WHB) domain were difficult to determine.…”

Section: Cryo-em Structures Of the Csn-crl2~n8 Complexmentioning

confidence: 87%

“…Electron microscopy (EM) based structural analysis has provided important insights into the mechanism of CRL1 regulation by CSN 16,19 . CRL4A and CRL3 have also been observed to form such complexes 20 .…”

Section: Discussionmentioning

confidence: 99%

“…CRL4A and CRL3 have also been observed to form such complexes 20 . However, despite intense interest, structural information of the CSN bound to CRLs remains limited to CRL1 12,16,19 , CRL4A 20 , and a low-resolution map of a dimeric CSN-CRL3-N8 20 complex. In particular the analysis of the CSN-CRL4A complex 20 identified at least three major steps by which CRL~N8 is deneddylated by the CSN.…”

Section: Discussionmentioning

confidence: 99%

“…In particular the analysis of the CSN-CRL4A complex 20 identified at least three major steps by which CRL~N8 is deneddylated by the CSN. In the first step, the extended N-terminal helical modules of CSN2 and CSN4 conformationally "clamp" the C-terminal domain of the CRL4A~N8 and RBX1 16,19,20 . The second step involves the release and consequent relocation of CSN5/CSN6 closer to NEDD8, brought about by disruption of the CSN4/CSN6 interface 20 .…”

Section: Discussionmentioning

confidence: 99%

“…S1-S2). The mutation H138A in the catalytic site of CSN5 subunit makes it possible to assemble CSN-CRL2~N8 complexes in which NEDD8 remains covalently attached over the time scale for experimentation 16,19 . This mutant form of CSN was used throughout the work described below, unless otherwise specified.…”

Section: Cryo-em Structures Of the Csn-crl2~n8 Complexmentioning

confidence: 99%

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Structural basis of Cullin-2 RING E3 ligase regulation by the COP9 signalosome

Faull

Lau

Martens

et al. 2018

Preprint

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MAINCullin-RING Ligases (CRLs) are modular, multi-subunit complexes that constitute a major class of ubiquitin E3 ligases 1,2 . CRLs coordinate the ubiquitination of substrates as either a signal for degradation via the 26S proteasome, or to alter the function of the target protein 2,3 . The CRL2 E3 ligase consists of a Cullin-2 (CUL2) scaffold in association with a catalytic RING-box protein (RBX1), with the substrate adaptors Elongin B (ELOB) and C (ELOC) at its N-terminal 4 . When associated with the von Hippel-Lindau (VHL) tumour suppressor substrate receptor, the CRL2 complex is the primary regulator of the Hypoxia Inducible Factor 1-α (HIF-1α) transcription factor 5,6 . Mutations in the interface between VHL, ELOB and ELOC can deactivate CRL2leading to an accumulation of HIF-1α, which can in turn drive tumorigenesis through the over-activation of oncogenes 7 . Moreover, CRL2 has recently been identified as a potential target for small molecular inhibitors and PROTACs -a new class of cancer drugs that promote degradation of tumorigenic gene products 8-10 . These fascinating systems have been described in detail by a number of excellent reviews 1,2,3 .Activation of CRL2, in common with other members of the CRL family, involves a cascade of E1, E2 and E3 enzymes, which conjugate the ubiquitin-like protein NEDD8 (N8) to residue K689 on the Cullin-2 scaffold 11 . In its activated state, CRL2~N8 (the ~ stylization denotes a covalent interaction) recruits the ubiquitin-conjugated E2 enzyme via the RING domain of RBX1 12 . Ubiquitination now takes place, covalently adding ubiquitin to the substrate molecule docked at the CRL2 N-terminal. The activity of CRL2 is negatively regulated by the 331 kDa Constitutive Photomorphogenesis 9 Signalosome (CSN) complex, frequently referred to as the COP9 signalosome complex [13][14][15] . The CSN was originally identified as consisting of eight subunits

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“…In our EM maps and the other published CSN-CRL structures 16,19,20 , the exact position of NEDD8 and the Cullin-2 Winged-Helix B (WHB) domain were difficult to determine.…”

Section: Cryo-em Structures Of the Csn-crl2~n8 Complexmentioning

confidence: 87%