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PMID: 10733526 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The KEN box: an APC recognition signal distinct from the D box targeted by Cdh1.

Genes & development ·Vol. 14 ·No. 6 ·2000-03-15 ·Pages 655-65

Pfleger CM, Kirschner MW

Abstract

The ordered progression through the cell cycle depends on regulating the abundance of several proteins through ubiquitin-mediated proteolysis. Degradation is precisely timed and specific. One key component of the degradation system, the anaphase promoting complex (APC), is a ubiquitin protein ligase. It is activated both during mitosis and late in mitosis/G(1), by the WD repeat proteins Cdc20 and Cdh1, respectively. These activators target distinct sets of substrates. Cdc20-APC requires a well-defined destruction box (D box), whereas Cdh1-APC confers a different and as yet unidentified specificity. We have determined the sequence specificity for Cdh1-APC using two assays, ubiquitination in a completely defined and purified system and degradation promoted by Cdh1-APC in Xenopus extracts. Cdc20 is itself a Cdh1-APC substrate. Vertebrate Cdc20 lacks a D box and therefore is recognized by Cdh1-APC through a different sequence. By analysis of Cdc20 as a substrate, we have identified a new recognition signal. This signal, composed of K-E-N, serves as a general targeting signal for Cdh1-APC. Like the D box, it is transposable to other proteins. Using the KEN box as a template, we have identified cell cycle genes Nek2 and B99 as additional Cdh1-APC substrates. Mutation in the KEN box stabilizes all three proteins against ubiquitination and degradation.

MeSH Terms
Amino Acid Sequence Anaphase-Promoting Complex-Cyclosome Animals Binding Sites Cdc20 Proteins Cdh1 Proteins Cell Cycle Proteins/metabolism Cyclin B/chemistry,metabolism Fungal Proteins/metabolism Ligases/genetics,metabolism Molecular Sequence Data Protein Sorting Signals/metabolism Saccharomyces cerevisiae Proteins Ubiquitin-Protein Ligase Complexes Ubiquitin-Protein Ligases Xenopus
Chemicals
CDC20 protein, S cerevisiae CDH1 protein, S cerevisiae Cdc20 Proteins Cdh1 Proteins Cell Cycle Proteins Cyclin B Fungal Proteins Protein Sorting Signals Saccharomyces cerevisiae Proteins Ubiquitin-Protein Ligase Complexes Anaphase-Promoting Complex-Cyclosome Ubiquitin-Protein Ligases Ligases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Pfleger C M
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115 USA.
Kirschner M W
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
Published
2000-03-15
Pages
655-65
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC316466
Subset
IM
Grants
NIGMS NIH HHS · R01 GM026875 · United States
NIGMS NIH HHS · R01 GM039023 · United States
NIGMS NIH HHS · GM26875 · United States
NIGMS NIH HHS · GM39023 · United States
External Links
PubMed source PMC Full Text View journal details
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