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

DOI: 10.1038/s41467-019-11772-y DOI Help

Authors: Sarah V. Faull (The Institute of Cancer Research) , Andy M. C. Lau (King’s College London) , Chloe Martens (King’s College London) , Zainab Ahdash (King’s College London) , Kjetil Hansen (King’s College London) , Hugo Yebenes (The Institute of Cancer Research; Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas) , Carla Schmidt (Martin Luther University Halle-Wittenberg) , Fabienne Beuron (The Institute of Cancer Research) , Nora B. Cronin (The Institute of Cancer Research) , Edward Morris (The Institute of Cancer Research) , Argyris Politis (King’s College London)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 10

State: Published (Approved)
Published: September 2019
Diamond Proposal Number(s): 15621 , 16023

Open Access Open Access

Abstract: Cullin-Ring E3 Ligases (CRLs) regulate a multitude of cellular pathways through specific substrate receptors. The COP9 signalosome (CSN) deactivates CRLs by removing NEDD8 from activated Cullins. Here we present structures of the neddylated and deneddylated CSN-CRL2 complexes by combining single-particle cryo-electron microscopy (cryo-EM) with chemical cross-linking mass spectrometry (XL-MS). These structures suggest a conserved mechanism of CSN activation, consisting of conformational clamping of the CRL2 substrate by CSN2/CSN4, release of the catalytic CSN5/CSN6 heterodimer and finally activation of the CSN5 deneddylation machinery. Using hydrogen-deuterium exchange (HDX)-MS we show that CRL2 activates CSN5/CSN6 in a neddylation-independent manner. The presence of NEDD8 is required to activate the CSN5 active site. Overall, by synergising cryo-EM with MS, we identify sensory regions of the CSN that mediate its stepwise activation and provide a framework for understanding the regulatory mechanism of other Cullin family members.

Journal Keywords: Cryoelectron microscopy; Enzyme mechanisms; Mass spectrometry; Neddylation

Subject Areas: Biology and Bio-materials, Medicine

Diamond Offline Facilities: Electron Bio-Imaging Centre (eBIC)
Instruments: Krios I-Titan Krios I at Diamond

Added On: 12/09/2019 15:42

Documents:
s41467-019-11772-y.pdf

Discipline Tags:

Life Sciences & Biotech Health & Wellbeing Cancer Drug Discovery Non-Communicable Diseases Structural biology

Technical Tags:

Microscopy Electron Microscopy (EM) Cryo Electron Microscopy (Cryo EM)