Autoinhibitory regulation of SCF-mediated ubiquitination by human cullin 1's C-terminal tail.
SCF (Skp1 x CUL1 x F-box protein x ROC1) E3 ubiquitin ligase and Cdc34 E2-conjugating enzyme catalyze polyubiquitination in a precisely regulated fashion. Here, we describe biochemical evidence suggesting an autoinhibitory role played by the human CUL1 ECTD (extreme C-terminal domain; spanning the C-terminal 50 amino acids), a region that is predicted to contact the ROC1 RING finger protein by structural studies. We showed that ECTD did not contribute to CUL1's stable association with ROC1. Remarkably, deletion of ECTD, or missense mutations designed to disrupt the predicted ECTD x ROC1 interaction, markedly increased the ability of SCF(betaTrCP2) to promote IkappaB alpha polyubiquitination and polyubiquitin chain assembly by Cdc34 in vitro. Thus, disruption of ECTD yields in vitro effects that parallel SCF activation by Nedd8 conjugation to CUL1. We propose that SCF may be subject to autoinhibitory regulation, in which Nedd8 conjugation acts as a molecular switch to drive the E3 into an active state by diminishing the inhibitory ECTD x ROC1 interaction.
Yamoah
Kosj
K
Department of Oncological Sciences, Mount Sinai School of Medicine, New York, NY 10029-6574, USA.
Oashi
Taiji
T
Sarikas
Antonio
A
Gazdoiu
Stefan
S
Osman
Roman
R
Pan
Zhen-Qiang
ZQ
D064173:Anaphase-Promoting Complex-Cyclosome; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism*; D044842:Cullin Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D006706:Homeostasis; D006801:Humans; D000075687:NEDD8 Protein; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D044763:Ubiquitin-Conjugating Enzymes; D043743:Ubiquitin-Protein Ligase Complexes; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D054875:Ubiquitination; D014452:Ubiquitins / Q000378:metabolism*
D002352:Carrier Proteins; C100318:Cullin 1; D044842:Cullin Proteins; D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; C479529:RBX1 protein, human; D014452:Ubiquitins; C577447:CDC34 protein, human; D044763:Ubiquitin-Conjugating Enzymes; D043743:Ubiquitin-Protein Ligase Complexes; D064173:Anaphase-Promoting Complex-Cyclosome; D044767:Ubiquitin-Protein Ligases