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

Sarah V Faull; Andy M C Lau; Chloe Martens; Zainab Ahdash; Kjetil Hansen; Hugo Yebenes; Carla Schmidt; Fabienne Beuron; Nora B Cronin; Edward P Morris; Argyris Politis

doi:10.1038/s41467-019-11772-y

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

Nat Commun. 2019 Aug 23;10(1):3814. doi: 10.1038/s41467-019-11772-y.

Authors

Affiliations

¹ Division of Structural Biology, The Institute of Cancer Research, London, SW3 6JB, UK.
² Department of Chemistry, King's College London, 7 Trinity Street, London, SE1 1DB, UK.
³ Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
⁴ Interdisciplinary Research Center HALOmem, Charles Tanford Protein Centre, Martin Luther University Halle-Wittenberg, Kurt-Mothes-Strasse 3a, 06120, Halle/Saale, Germany.
⁵ Division of Structural Biology, The Institute of Cancer Research, London, SW3 6JB, UK. ed.morris@icr.ac.uk.
⁶ Department of Chemistry, King's College London, 7 Trinity Street, London, SE1 1DB, UK. argyris.politis@kcl.ac.uk.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adaptor Proteins, Signal Transducing / isolation & purification
Adaptor Proteins, Signal Transducing / metabolism
Animals
COP9 Signalosome Complex / isolation & purification
COP9 Signalosome Complex / metabolism
COP9 Signalosome Complex / ultrastructure*
Cryoelectron Microscopy
Intracellular Signaling Peptides and Proteins / isolation & purification
Intracellular Signaling Peptides and Proteins / metabolism
Mass Spectrometry
NEDD8 Protein / isolation & purification
NEDD8 Protein / metabolism
NEDD8 Protein / ultrastructure*
Peptide Hydrolases / isolation & purification
Peptide Hydrolases / metabolism
Peptide Hydrolases / ultrastructure*
Protein Processing, Post-Translational
Recombinant Proteins / isolation & purification
Recombinant Proteins / metabolism
Recombinant Proteins / ultrastructure
Sf9 Cells
Ubiquitin-Protein Ligases / isolation & purification
Ubiquitin-Protein Ligases / metabolism
Ubiquitin-Protein Ligases / ultrastructure*

Substances

Adaptor Proteins, Signal Transducing
COPS6 protein, human
Intracellular Signaling Peptides and Proteins
NEDD8 Protein
NEDD8 protein, human
Recombinant Proteins
CULL-RING ligase, human
Ubiquitin-Protein Ligases
Peptide Hydrolases
COPS5 protein, human
COP9 Signalosome Complex

Abstract

Publication types

MeSH terms

Substances

Grants and funding