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PMID: 11850407 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Pseudosubstrate regulation of the SCF(beta-TrCP) ubiquitin ligase by hnRNP-U.

Genes & development ·Vol. 16 ·No. 4 ·2002-02-15 ·Pages 439-51

Davis M, Hatzubai A, Andersen JS, Ben-Shushan E, Fisher GZ, Yaron A, Bauskin A, Mercurio F, Mann M, Ben-Neriah Y

Abstract

beta-TrCP/E3RS (E3RS) is the F-box protein that functions as the receptor subunit of the SCF(beta-TrCP) ubiquitin ligase (E3). Surprisingly, although its two recognized substrates, IkappaB(alpha) and beta-catenin, are present in the cytoplasm, we have found that E3RS is located predominantly in the nucleus. Here we report the isolation of the major E3RS-associated protein, hnRNP-U, an abundant nuclear phosphoprotein. This protein occupies E3RS in a specific and stoichiometric manner, stabilizes the E3 component, and is likely responsible for its nuclear localization. hnRNP-U binding was abolished by competition with a pIkappaB(alpha) peptide, or by a specific point mutation in the E3RS WD region, indicating an E3-substrate-type interaction. However, unlike pI(kappa)Balpha, which is targeted by SCF(beta-TrCP) for degradation, the E3-bound hnRNP-U is stable and is, therefore, a pseudosubstrate. Consequently, hnRNP-U engages a highly neddylated active SCF(beta-TrCP), which dissociates in the presence of a high-affinity substrate, resulting in ubiquitination of the latter. Our study points to a novel regulatory mechanism, which secures the localization, stability, substrate binding threshold, and efficacy of a specific protein-ubiquitin ligase.

MeSH Terms
Active Transport, Cell Nucleus Animals CHO Cells Cricetinae Cricetulus DNA-Binding Proteins/metabolism,pharmacology GTP-Binding Proteins/metabolism HeLa Cells Heterogeneous-Nuclear Ribonucleoprotein U Heterogeneous-Nuclear Ribonucleoproteins Humans I-kappa B Proteins Macromolecular Substances Mice Models, Biological NF-KappaB Inhibitor alpha Nuclear Proteins/metabolism Peptide Fragments/metabolism,pharmacology Peptide Synthases/metabolism Point Mutation Protein Binding Protein Interaction Mapping Protein Processing, Post-Translational/physiology Ribonucleoproteins/physiology SKP Cullin F-Box Protein Ligases Subcellular Fractions/metabolism Substrate Specificity Ubiquitin/metabolism beta-Transducin Repeat-Containing Proteins
Chemicals
BTRC protein, human Btrc protein, mouse DNA-Binding Proteins Heterogeneous-Nuclear Ribonucleoprotein U Heterogeneous-Nuclear Ribonucleoproteins I-kappa B Proteins Macromolecular Substances NFKBIA protein, human Nfkbia protein, mouse Nuclear Proteins Peptide Fragments Ribonucleoproteins Ubiquitin beta-Transducin Repeat-Containing Proteins NF-KappaB Inhibitor alpha SKP Cullin F-Box Protein Ligases GTP-Binding Proteins Peptide Synthases
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Davis Matti
The Lautenberg Center for Immunology, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
Hatzubai Ada
Andersen Jens S
Ben-Shushan Etti
Fisher Gregory Zvi
Yaron Avraham
Bauskin Asne
Mercurio Frank
Mann Matthias
Ben-Neriah Yinon
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.218702
Published
2002-02-15
Pages
439-51
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC155337
Subset
IM
External Links
PubMed source DOI: 10.1101/gad.218702 PMC Full Text View journal details
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