TY - JOUR
T1 - Structural basis of Cullin 2 RING E3 ligase regulation by the COP9 signalosome
AU - Faull, Sarah V
AU - Lau, Andy
AU - Martens, Chloe Paule Vinciane Julienne
AU - Ahdash, Zainab Mohamed
AU - Hansen, Kjetil
AU - Yebenes, Hugo
AU - Schmidt, Carla
AU - Beuron, Fabienne
AU - Cronin, Nora
AU - Morris, Edward P.
AU - Politis, Anargyros
PY - 2019/12/1
Y1 - 2019/12/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85071300293&partnerID=8YFLogxK
U2 - 10.1038/s41467-019-11772-y
DO - 10.1038/s41467-019-11772-y
M3 - Article
SN - 2041-1723
VL - 10
SP - 1
EP - 13
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 3814
ER -