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PMID: 10385629 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cdc53/cullin and the essential Hrt1 RING-H2 subunit of SCF define a ubiquitin ligase module that activates the E2 enzyme Cdc34.

Genes & development ·Vol. 13 ·No. 12 ·1999-06-15 ·Pages 1614-26

Seol JH, Feldman RM, Zachariae W, Shevchenko A, Correll CC, Lyapina S, Chi Y, Galova M, Claypool J, Sandmeyer S, Nasmyth K, Deshaies RJ, Shevchenko A, Deshaies RJ

Abstract

SCFCdc4 (Skp1, Cdc53/cullin, F-box protein) defines a family of modular ubiquitin ligases (E3s) that regulate diverse processes including cell cycle, immune response, and development. Mass spectrometric analysis of proteins copurifying with Cdc53 identified the RING-H2 finger protein Hrt1 as a subunit of SCF. Hrt1 shows striking similarity to the Apc11 subunit of anaphase-promoting complex. Conditional inactivation of hrt1(ts) results in stabilization of the SCFCdc4 substrates Sic1 and Cln2 and cell cycle arrest at G1/S. Hrt1 assembles into recombinant SCF complexes and individually binds Cdc4, Cdc53 and Cdc34, but not Skp1. Hrt1 stimulates the E3 activity of recombinant SCF potently and enables the reconstitution of Cln2 ubiquitination by recombinant SCFGrr1. Surprisingly, SCF and the Cdc53/Hrt1 subcomplex activate autoubiquitination of Cdc34 E2 enzyme by a mechanism that does not appear to require a reactive thiol. The highly conserved human HRT1 complements the lethality of hrt1Delta, and human HRT2 binds CUL-1. We conclude that Cdc53/Hrt1 comprise a highly conserved module that serves as the functional core of a broad variety of heteromeric ubiquitin ligases.

MeSH Terms
Amino Acid Sequence Anaphase-Promoting Complex-Cyclosome Animals Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome Basic Helix-Loop-Helix Transcription Factors Binding Sites Cell Cycle Proteins/genetics,isolation & purification,metabolism Cullin Proteins Enzyme Activation F-Box Proteins F-Box-WD Repeat-Containing Protein 7 Humans Ligases/metabolism Molecular Sequence Data Recombinant Fusion Proteins/metabolism S-Phase Kinase-Associated Proteins SKP Cullin F-Box Protein Ligases Saccharomyces cerevisiae Proteins Tripeptidyl-Peptidase 1 Ubiquitin-Conjugating Enzymes Ubiquitin-Protein Ligase Complexes Ubiquitin-Protein Ligases Ubiquitins/metabolism
Chemicals
Basic Helix-Loop-Helix Transcription Factors Cdc53 protein, S cerevisiae Cell Cycle Proteins Cullin Proteins F-Box Proteins F-Box-WD Repeat-Containing Protein 7 FBXW7 protein, human HEY1 protein, human Recombinant Fusion Proteins S-Phase Kinase-Associated Proteins Saccharomyces cerevisiae Proteins Tripeptidyl-Peptidase 1 Ubiquitins CDC34 protein, S cerevisiae CDC34 protein, human Ubiquitin-Conjugating Enzymes Ubiquitin-Protein Ligase Complexes ANAPC11 protein, human Anaphase-Promoting Complex-Cyclosome Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome HRT1 protein, S cerevisiae SKP Cullin F-Box Protein Ligases Ubiquitin-Protein Ligases TPP1 protein, human Ligases
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Seol J H
Division of Biology, California Institute of Technology, Pasadena, California 91125, USA.
Feldman R M
Zachariae W
Shevchenko A
Correll C C
Lyapina S
Chi Y
Galova M
Claypool J
Sandmeyer S
Nasmyth K
Deshaies R J
Shevchenko A
Deshaies R J
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.13.12.1614
Published
1999-06-15
Pages
1614-26
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC316801
Subset
IM
Grants
Wellcome Trust · United Kingdom
NIGMS NIH HHS · GM52466-01 · United States
External Links
PubMed source DOI: 10.1101/gad.13.12.1614 PMC Full Text View journal details
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