Home LiteratureArticle Details
PMID: 10713156 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Nedd8 modification of cul-1 activates SCF(beta(TrCP))-dependent ubiquitination of IkappaBalpha.

Molecular and cellular biology ·Vol. 20 ·No. 7 ·2000-04-00 ·Pages 2326-33

Read MA, Brownell JE, Gladysheva TB, Hottelet M, Parent LA, Coggins MB, Pierce JW, Podust VN, Luo RS, Chau V, Palombella VJ

Abstract

Regulation of NF-kappaB occurs through phosphorylation-dependent ubiquitination of IkappaBalpha, which is degraded by the 26S proteasome. Recent studies have shown that ubiquitination of IkappaBalpha is carried out by a ubiquitin-ligase enzyme complex called SCF(beta(TrCP)). Here we show that Nedd8 modification of the Cul-1 component of SCF(beta(TrCP)) is important for function of SCF(beta(TrCP)) in ubiquitination of IkappaBalpha. In cells, Nedd8-conjugated Cul-1 was complexed with two substrates of SCF(beta(TrCP)), phosphorylated IkappaBalpha and beta-catenin, indicating that Nedd8-Cul-1 conjugates are part of SCF(beta(TrCP)) in vivo. Although only a minute fraction of total cellular Cul-1 is modified by Nedd8, the Cul-1 associated with ectopically expressed betaTrCP was highly enriched for the Nedd8-conjugated form. Moreover, optimal ubiquitination of IkappaBalpha required Nedd8 and the Nedd8-conjugating enzyme, Ubc12. The site of Nedd8 ligation to Cul-1 is essential, as SCF(beta(TrCP)) containing a K720R mutant of Cul-1 only weakly supported IkappaBalpha ubiquitination compared to SCF(beta(TrCP)) containing WT Cul-1, suggesting that the Nedd8 ligation of Cul-1 affects the ubiquitination activity of SCF(beta(TrCP)). These observations provide a functional link between the highly related ubiquitin and Nedd8 pathways of protein modification and show how they operate together to selectively target the signal-dependent degradation of IkappaBalpha.

MeSH Terms
Amino Acid Sequence Cell Cycle Proteins Cell Line Cullin Proteins Cytoskeletal Proteins/metabolism DNA-Binding Proteins/metabolism GTP-Binding Proteins/genetics,metabolism Helminth Proteins/genetics,metabolism Humans I-kappa B Proteins Kinetics Molecular Sequence Data Multienzyme Complexes/metabolism NEDD8 Protein Peptide Synthases/metabolism Phosphorylation SKP Cullin F-Box Protein Ligases Saccharomyces cerevisiae Proteins Sequence Alignment Trans-Activators Transfection Ubiquitins/metabolism beta Catenin beta-Transducin Repeat-Containing Proteins
Chemicals
BTRC protein, human CTNNB1 protein, human Cell Cycle Proteins Cullin 1 Cullin Proteins Cytoskeletal Proteins DNA-Binding Proteins Helminth Proteins I kappa B beta protein I-kappa B Proteins Multienzyme Complexes NEDD8 Protein NEDD8 protein, human Saccharomyces cerevisiae Proteins Trans-Activators UBC12 protein, S cerevisiae Ubiquitins beta Catenin beta-Transducin Repeat-Containing Proteins SKP Cullin F-Box Protein Ligases GTP-Binding Proteins Peptide Synthases
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Read M A
LeukoSite, Inc., Cambridge, Massachusetts 02139, USA. [email protected]
Brownell J E
Gladysheva T B
Hottelet M
Parent L A
Coggins M B
Pierce J W
Podust V N
Luo R S
Chau V
Palombella V J
References (64)
64 references, click to expand
  1. Cdc53 is a scaffold protein for multiple Cdc34/Skp1/F-box proteincomplexes that regulate cell division and methionine biosynthesis in yeast.
    Genes Dev. 1998 Mar 1;12(5):692-705 PMID: 9499404
  2. Inhibition of NF-kappa-B cellular function via specific targeting of the I-kappa-B-ubiquitin ligase.
    EMBO J. 1997 Nov 3;16(21):6486-94 PMID: 9351830
  3. 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
  4. Combinatorial control in ubiquitin-dependent proteolysis: don't Skp the F-box hypothesis.
    Trends Genet. 1998 Jun;14(6):236-43 PMID: 9635407
  5. A novel human WD protein, h-beta TrCp, that interacts with HIV-1 Vpu connects CD4 to the ER degradation pathway through an F-box motif.
    Mol Cell. 1998 Mar;1(4):565-74 PMID: 9660940
  6. Human CUL-1, but not other cullin family members, selectively interacts with SKP1 to form a complex with SKP2 and cyclin A.
    Cell Growth Differ. 1998 Jun;9(6):435-49 PMID: 9663463
  7. MEKK1 activates both IkappaB kinase alpha and IkappaB kinase beta.
    Proc Natl Acad Sci U S A. 1998 Aug 4;95(16):9319-24 PMID: 9689078
  8. A new NEDD8-ligating system for cullin-4A.
    Genes Dev. 1998 Aug 1;12(15):2263-8 PMID: 9694792
  9. Identification of the receptor component of the IkappaBalpha-ubiquitin ligase.
    Nature. 1998 Dec 10;396(6711):590-4 PMID: 9859996
  10. The ubiquitin-proteasome pathway: on protein death and cell life.
    EMBO J. 1998 Dec 15;17(24):7151-60 PMID: 9857172
  11. Structure of an IkappaBalpha/NF-kappaB complex.
    Cell. 1998 Dec 11;95(6):749-58 PMID: 9865693
  12. Inhibition of NF-kappa B activation in vitro and in vivo: role of 26S proteasome.
    Methods Enzymol. 1999;300:345-63 PMID: 9919536
  13. 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
  14. Signal-induced ubiquitination of IkappaBalpha by the F-box protein Slimb/beta-TrCP.
    Genes Dev. 1999 Feb 1;13(3):284-94 PMID: 9990853
  15. The human F box protein beta-Trcp associates with the Cul1/Skp1 complex and regulates the stability of beta-catenin.
    Oncogene. 1999 Jan 28;18(4):849-54 PMID: 10023660
  16. IkappaBalpha ubiquitination is catalyzed by an SCF-like complex containing Skp1, cullin-1, and two F-box/WD40-repeat proteins, betaTrCP1 and betaTrCP2.
    Biochem Biophys Res Commun. 1999 Mar 5;256(1):127-32 PMID: 10066435
  17. 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
  18. Inducible degradation of IkappaBalpha by the proteasome requires interaction with the F-box protein h-betaTrCP.
    J Biol Chem. 1999 Mar 19;274(12):7941-5 PMID: 10075690
  19. The F-box protein beta-TrCP associates with phosphorylated beta-catenin and regulates its activity in the cell.
    Curr Biol. 1999 Feb 25;9(4):207-10 PMID: 10074433
  20. Identification of NEDD8-conjugation site in human cullin-2.
    Biochem Biophys Res Commun. 1999 Apr 2;257(1):100-5 PMID: 10092517
  21. Ubiquitin-dependent degradation of IkappaBalpha is mediated by a ubiquitin ligase Skp1/Cul 1/F-box protein FWD1.
    Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3859-63 PMID: 10097128
  22. Identification of the activating and conjugating enzymes of the NEDD8 conjugation pathway.
    J Biol Chem. 1999 Apr 23;274(17):12036-42 PMID: 10207026
  23. Rbx1, a component of the VHL tumor suppressor complex and SCF ubiquitin ligase.
    Science. 1999 Apr 23;284(5414):657-61 PMID: 10213691
  24. Reconstitution of G1 cyclin ubiquitination with complexes containing SCFGrr1 and Rbx1.
    Science. 1999 Apr 23;284(5414):662-5 PMID: 10213692
  25. Recruitment of a ROC1-CUL1 ubiquitin ligase by Skp1 and HOS to catalyze the ubiquitination of I kappa B alpha.
    Mol Cell. 1999 Apr;3(4):527-33 PMID: 10230406
  26. 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
  27. One ring to rule a superfamily of E3 ubiquitin ligases.
    Science. 1999 Apr 23;284(5414):601, 603-4 PMID: 10328744
  28. HOS, a human homolog of Slimb, forms an SCF complex with Skp1 and Cullin1 and targets the phosphorylation-dependent degradation of IkappaB and beta-catenin.
    Oncogene. 1999 Mar 25;18(12):2039-46 PMID: 10321728
  29. Conjugation of the ubiquitin-like protein NEDD8 to cullin-2 is linked to von Hippel-Lindau tumor suppressor function.
    Proc Natl Acad Sci U S A. 1999 May 11;96(10):5510-5 PMID: 10318914
  30. Identification of the ubiquitin carrier proteins, E2s, involved in signal-induced conjugation and subsequent degradation of IkappaBalpha.
    J Biol Chem. 1999 May 21;274(21):14823-30 PMID: 10329681
  31. beta-Trcp couples beta-catenin phosphorylation-degradation and regulates Xenopus axis formation.
    Proc Natl Acad Sci U S A. 1999 May 25;96(11):6273-8 PMID: 10339577
  32. 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
  33. Identification of an SCF ubiquitin-ligase complex required for auxin response in Arabidopsis thaliana.
    Genes Dev. 1999 Jul 1;13(13):1678-91 PMID: 10398681
  34. 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
  35. 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
  36. The von Hippel-Lindau tumor suppressor gene product promotes, but is not essential for, NEDD8 conjugation to cullin-2.
    J Biol Chem. 1999 Dec 10;274(50):36025-9 PMID: 10585493
  37. Components of ubiquitin-protein ligase system. Resolution, affinity purification, and role in protein breakdown.
    J Biol Chem. 1983 Jul 10;258(13):8206-14 PMID: 6305978
  38. Ubiquitin-aldehyde: a general inhibitor of ubiquitin-recycling processes.
    Proc Natl Acad Sci U S A. 1987 Apr;84(7):1829-33 PMID: 3031653
  39. Identification of a set of genes with developmentally down-regulated expression in the mouse brain.
    Biochem Biophys Res Commun. 1992 Jun 30;185(3):1155-61 PMID: 1378265
  40. Structure of a fibronectin type III domain from tenascin phased by MAD analysis of the selenomethionyl protein.
    Science. 1992 Nov 6;258(5084):987-91 PMID: 1279805
  41. The ubiquitin-proteasome pathway is required for processing the NF-kappa B1 precursor protein and the activation of NF-kappa B.
    Cell. 1994 Sep 9;78(5):773-85 PMID: 8087845
  42. Identification of a human ubiquitin-conjugating enzyme that mediates the E6-AP-dependent ubiquitination of p53.
    Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8797-801 PMID: 8090726
  43. Signal-induced site-specific phosphorylation targets I kappa B alpha to the ubiquitin-proteasome pathway.
    Genes Dev. 1995 Jul 1;9(13):1586-97 PMID: 7628694
  44. Stimulation-dependent I kappa B alpha phosphorylation marks the NF-kappa B inhibitor for degradation via the ubiquitin-proteasome pathway.
    Proc Natl Acad Sci U S A. 1995 Nov 7;92(23):10599-603 PMID: 7479848
  45. Identification of a family of closely related human ubiquitin conjugating enzymes.
    J Biol Chem. 1995 Dec 22;270(51):30408-14 PMID: 8530467
  46. The NF-kappa B and I kappa B proteins: new discoveries and insights.
    Annu Rev Immunol. 1996;14:649-83 PMID: 8717528
  47. SKP1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif, the F-box.
    Cell. 1996 Jul 26;86(2):263-74 PMID: 8706131
  48. 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
  49. Kinetic studies on the inhibition of isopeptidase T by ubiquitin aldehyde.
    Biochemistry. 1996 Oct 1;35(39):12893-900 PMID: 8841133
  50. NF-kappa B: ten years after.
    Cell. 1996 Oct 4;87(1):13-20 PMID: 8858144
  51. An essential role for NF-kappaB in preventing TNF-alpha-induced cell death.
    Science. 1996 Nov 1;274(5288):782-4 PMID: 8864118
  52. TNF- and cancer therapy-induced apoptosis: potentiation by inhibition of NF-kappaB.
    Science. 1996 Nov 1;274(5288):784-7 PMID: 8864119
  53. Suppression of TNF-alpha-induced apoptosis by NF-kappaB.
    Science. 1996 Nov 1;274(5288):787-9 PMID: 8864120
  54. Dissection of TNF receptor 1 effector functions: JNK activation is not linked to apoptosis while NF-kappaB activation prevents cell death.
    Cell. 1996 Nov 1;87(3):565-76 PMID: 8898208
  55. Nuclear factor-kappaB: a pivotal transcription factor in chronic inflammatory diseases.
    N Engl J Med. 1997 Apr 10;336(15):1066-71 PMID: 9091804
  56. Dominant-negative cyclin-selective ubiquitin carrier protein E2-C/UbcH10 blocks cells in metaphase.
    Proc Natl Acad Sci U S A. 1997 Mar 18;94(6):2362-7 PMID: 9122200
  57. beta-catenin is a target for the ubiquitin-proteasome pathway.
    EMBO J. 1997 Jul 1;16(13):3797-804 PMID: 9233789
  58. Signal transduction. IkappaB kinase all zipped up.
    Nature. 1997 Aug 7;388(6642):519-21 PMID: 9252183
  59. F-box proteins are receptors that recruit phosphorylated substrates to the SCF ubiquitin-ligase complex.
    Cell. 1997 Oct 17;91(2):209-19 PMID: 9346238
  60. A complex of Cdc4p, Skp1p, and Cdc53p/cullin catalyzes ubiquitination of the phosphorylated CDK inhibitor Sic1p.
    Cell. 1997 Oct 17;91(2):221-30 PMID: 9346239
  61. Characterization of NEDD8, a developmentally down-regulated ubiquitin-like protein.
    J Biol Chem. 1997 Nov 7;272(45):28557-62 PMID: 9353319
  62. NF-kappaB activation: the I kappaB kinase revealed?
    Cell. 1997 Oct 31;91(3):299-302 PMID: 9363938
  63. Catalysis by a multiprotein IkappaB kinase complex.
    Science. 1997 Oct 31;278(5339):818-9 PMID: 9381193
  64. 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
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.20.7.2326-2333.2000
Email
[email protected]
Published
2000-04-00
Pages
2326-33
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC85397
Subset
IM
Grants
NIGMS NIH HHS · GM53136 · United States
External Links
PubMed source DOI: 10.1128/MCB.20.7.2326-2333.2000 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