title: NEDD8 recruits E2-ubiquitin to SCF E3 ligase.
abstract: NEDD8/Rub1 is a ubiquitin (Ub)-like post-translational modifier that is covalently linked to cullin (Cul)-family proteins in a manner analogous to ubiquitylation. NEDD8 is known to enhance the ubiquitylating activity of the SCF complex (composed of Skp1, Cul-1, ROC1 and F-box protein), but the mechanistic role is largely unknown. Using an in vitro reconstituted system, we report here that NEDD8 modification of Cul-1 enhances recruitment of Ub-conjugating enzyme Ubc4 (E2) to the SCF complex (E3). This recruitment requires thioester linkage of Ub to Ubc4. Our findings indicate that the NEDD8-modifying system accelerates the formation of the E2-E3 complex, which stimulates protein polyubiquitylation.
authors.affiliation: Department of Gastroenterology, Faculty of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Kawakami
authors.forename: T
authors.initials: T
authors.lastname: Chiba
authors.forename: T
authors.initials: T
authors.lastname: Suzuki
authors.forename: K
authors.initials: K
authors.lastname: Iwai
authors.forename: K
authors.initials: K
authors.lastname: Yamanaka
authors.forename: N
authors.initials: N
authors.lastname: Minato
authors.forename: H
authors.initials: H
authors.lastname: Suzuki
authors.forename: N
authors.initials: N
authors.lastname: Shimbara
authors.forename: Y
authors.initials: Y
authors.lastname: Hidaka
authors.forename: F
authors.initials: F
authors.lastname: Osaka
authors.forename: M
authors.initials: M
authors.lastname: Omata
authors.forename: K
authors.initials: K
authors.lastname: Tanaka
mesh_terms: D064173:Anaphase-Promoting Complex-Cyclosome; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D020672:I-kappa B Proteins; D008025:Ligases / Q000235:genetics / Q000378:metabolism*; D000075687:NEDD8 Protein; D000072000:NF-KappaB Inhibitor alpha; D010453:Peptide Synthases / Q000378:metabolism*; D044843:SKP Cullin F-Box Protein Ligases; D044763:Ubiquitin-Conjugating Enzymes; D043743:Ubiquitin-Protein Ligase Complexes; D044767:Ubiquitin-Protein Ligases; D014452:Ubiquitins / Q000378:metabolism / Q000502:physiology*
chemical_list: D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; C000606204:NFKBIA protein, human; D014452:Ubiquitins; D000072000:NF-KappaB Inhibitor alpha; D044763:Ubiquitin-Conjugating Enzymes; D043743:Ubiquitin-Protein Ligase Complexes; C063109:ubiquitin-conjugating enzyme UBC4; D064173:Anaphase-Promoting Complex-Cyclosome; D044843:SKP Cullin F-Box Protein Ligases; D044767:Ubiquitin-Protein Ligases; D008025:Ligases; D010453:Peptide Synthases