Home LiteratureArticle Details
PMID: 11483504 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

NEDD8 recruits E2-ubiquitin to SCF E3 ligase.

The EMBO journal ·Vol. 20 ·No. 15 ·2001-08-01 ·Pages 4003-12

Kawakami T, Chiba T, Suzuki T, Iwai K, Yamanaka K, Minato N, Suzuki H, Shimbara N, Hidaka Y, Osaka F, Omata M, Tanaka K

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.

MeSH Terms
Anaphase-Promoting Complex-Cyclosome Cell Line DNA-Binding Proteins/metabolism Humans I-kappa B Proteins Ligases/genetics,metabolism NEDD8 Protein NF-KappaB Inhibitor alpha Peptide Synthases/metabolism SKP Cullin F-Box Protein Ligases Ubiquitin-Conjugating Enzymes Ubiquitin-Protein Ligase Complexes Ubiquitin-Protein Ligases Ubiquitins/metabolism,physiology
Chemicals
DNA-Binding Proteins I-kappa B Proteins NEDD8 Protein NEDD8 protein, human NFKBIA protein, human Ubiquitins NF-KappaB Inhibitor alpha Ubiquitin-Conjugating Enzymes Ubiquitin-Protein Ligase Complexes ubiquitin-conjugating enzyme UBC4 Anaphase-Promoting Complex-Cyclosome SKP Cullin F-Box Protein Ligases Ubiquitin-Protein Ligases Ligases Peptide Synthases
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Kawakami T
Department of Gastroenterology, Faculty of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan.
Chiba T
Suzuki T
Iwai K
Yamanaka K
Minato N
Suzuki H
Shimbara N
Hidaka Y
Osaka F
Omata M
Tanaka K
References (40)
40 references, click to expand
  1. Mechanistic aspects of NF-kappa B regulation: the emerging role of phosphorylation and proteolysis.
    Immunity. 1995 Sep;3(3):263-72 PMID: 7552992
  2. ROC1, a homolog of APC11, represents a family of cullin partners with an associated ubiquitin ligase activity.
    Mol Cell. 1999 Apr;3(4):535-41 PMID: 10230407
  3. Cdc53/cullin and the essential Hrt1 RING-H2 subunit of SCF define a ubiquitin ligase module that activates the E2 enzyme Cdc34.
    Genes Dev. 1999 Jun 15;13(12):1614-26 PMID: 10385629
  4. SGT1 encodes an essential component of the yeast kinetochore assembly pathway and a novel subunit of the SCF ubiquitin ligase complex.
    Mol Cell. 1999 Jul;4(1):21-33 PMID: 10445024
  5. The Rbx1 subunit of SCF and VHL E3 ubiquitin ligase activates Rub1 modification of cullins Cdc53 and Cul2.
    Genes Dev. 1999 Nov 15;13(22):2928-33 PMID: 10579999
  6. Covalent modification of all members of human cullin family proteins by NEDD8.
    Oncogene. 1999 Nov 18;18(48):6829-34 PMID: 10597293
  7. SCF and Cullin/Ring H2-based ubiquitin ligases.
    Annu Rev Cell Dev Biol. 1999;15:435-67 PMID: 10611969
  8. Homodimer of two F-box proteins betaTrCP1 or betaTrCP2 binds to IkappaBalpha for signal-dependent ubiquitination.
    J Biol Chem. 2000 Jan 28;275(4):2877-84 PMID: 10644755
  9. Function of the ubiquitin-proteasome pathway in auxin response.
    Trends Biochem Sci. 2000 Mar;25(3):133-8 PMID: 10694884
  10. Nedd8 modification of cul-1 activates SCF(beta(TrCP))-dependent ubiquitination of IkappaBalpha.
    Mol Cell Biol. 2000 Apr;20(7):2326-33 PMID: 10713156
  11. The amyloid precursor protein-binding protein APP-BP1 drives the cell cycle through the S-M checkpoint and causes apoptosis in neurons.
    J Biol Chem. 2000 Mar 24;275(12):8929-35 PMID: 10722740
  12. Modification of cullin-1 by ubiquitin-like protein Nedd8 enhances the activity of SCF(skp2) toward p27(kip1).
    Biochem Biophys Res Commun. 2000 Apr 21;270(3):1093-6 PMID: 10772955
  13. A Nedd8 conjugation pathway is essential for proteolytic targeting of p27Kip1 by ubiquitination.
    Proc Natl Acad Sci U S A. 2000 Apr 25;97(9):4579-84 PMID: 10781063
  14. Ubiquitin-like proteins: new wines in new bottles.
    Gene. 2000 May 2;248(1-2):1-14 PMID: 10806345
  15. The 26S proteasome: a molecular machine designed for controlled proteolysis.
    Annu Rev Biochem. 1999;68:1015-68 PMID: 10872471
  16. Covalent modifier NEDD8 is essential for SCF ubiquitin-ligase in fission yeast.
    EMBO J. 2000 Jul 3;19(13):3475-84 PMID: 10880460
  17. Ubiquitin and its kin: how close are the family ties?
    Trends Cell Biol. 2000 Aug;10(8):335-42 PMID: 10884686
  18. Evolution and function of ubiquitin-like protein-conjugation systems.
    Nat Cell Biol. 2000 Aug;2(8):E153-7 PMID: 10934491
  19. The CUL1 C-terminal sequence and ROC1 are required for efficient nuclear accumulation, NEDD8 modification, and ubiquitin ligase activity of CUL1.
    Mol Cell Biol. 2000 Nov;20(21):8185-97 PMID: 11027288
  20. Conjugation of Nedd8 to CUL1 enhances the ability of the ROC1-CUL1 complex to promote ubiquitin polymerization.
    J Biol Chem. 2000 Oct 13;275(41):32317-24 PMID: 10921923
  21. The F-box protein family.
    Genome Biol. 2000;1(5):REVIEWS3002 PMID: 11178263
  22. The cell-cycle regulatory protein Cks1 is required for SCF(Skp2)-mediated ubiquitinylation of p27.
    Nat Cell Biol. 2001 Mar;3(3):321-4 PMID: 11231585
  23. Reconstitution of p53-ubiquitinylation reactions from purified components: the role of human ubiquitin-conjugating enzyme UBC4 and E6-associated protein (E6AP).
    Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3264-8 PMID: 7724550
  24. Activation of the IkappaB alpha kinase complex by MEKK1, a kinase of the JNK pathway.
    Cell. 1997 Jan 24;88(2):213-22 PMID: 9008162
  25. Association of human CUL-1 and ubiquitin-conjugating enzyme CDC34 with the F-box protein p45(SKP2): evidence for evolutionary conservation in the subunit composition of the CDC34-SCF pathway.
    EMBO J. 1998 Jan 15;17(2):368-83 PMID: 9430629
  26. There's the rub: a novel ubiquitin-like modification linked to cell cycle regulation.
    Genes Dev. 1998 Apr 1;12(7):901-7 PMID: 9531529
  27. Modification of yeast Cdc53p by the ubiquitin-related protein rub1p affects function of the SCFCdc4 complex.
    Genes Dev. 1998 Apr 1;12(7):914-26 PMID: 9531531
  28. A novel protein modification pathway related to the ubiquitin system.
    EMBO J. 1998 Apr 15;17(8):2208-14 PMID: 9545234
  29. NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses.
    Annu Rev Immunol. 1998;16:225-60 PMID: 9597130
  30. The ubiquitin-related protein RUB1 and auxin response in Arabidopsis.
    Science. 1998 Jun 12;280(5370):1760-3 PMID: 9624055
  31. A new NEDD8-ligating system for cullin-4A.
    Genes Dev. 1998 Aug 1;12(15):2263-8 PMID: 9694792
  32. The ubiquitin system.
    Annu Rev Biochem. 1998;67:425-79 PMID: 9759494
  33. Identification of the receptor component of the IkappaBalpha-ubiquitin ligase.
    Nature. 1998 Dec 10;396(6711):590-4 PMID: 9859996
  34. The SCFbeta-TRCP-ubiquitin ligase complex associates specifically with phosphorylated destruction motifs in IkappaBalpha and beta-catenin and stimulates IkappaBalpha ubiquitination in vitro.
    Genes Dev. 1999 Feb 1;13(3):270-83 PMID: 9990852
  35. A ubiquitin ligase complex essential for the NF-kappaB, Wnt/Wingless, and Hedgehog signaling pathways.
    Genes Dev. 1999 Mar 1;13(5):505-10 PMID: 10072378
  36. A novel ubiquitination factor, E4, is involved in multiubiquitin chain assembly.
    Cell. 1999 Mar 5;96(5):635-44 PMID: 10089879
  37. Identification of the activating and conjugating enzymes of the NEDD8 conjugation pathway.
    J Biol Chem. 1999 Apr 23;274(17):12036-42 PMID: 10207026
  38. Rbx1, a component of the VHL tumor suppressor complex and SCF ubiquitin ligase.
    Science. 1999 Apr 23;284(5414):657-61 PMID: 10213691
  39. IkappaB kinases: kinsmen with different crafts.
    Science. 1999 Apr 9;284(5412):271-3 PMID: 10232975
  40. The NF-kappa B and I kappa B proteins: new discoveries and insights.
    Annu Rev Immunol. 1996;14:649-83 PMID: 8717528
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.15.4003
Published
2001-08-01
Pages
4003-12
Language
English
Region
England
NLM ID
8208664
PMCID
PMC149148
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.15.4003 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2