The cell-cycle regulatory protein Cks1 is required for SCFSkp2-mediated ubiquitinylation of p27

Dvora Ganoth, Gil Bornstein, Tun K. Ko, Brett Larsen, Mike Tyers, Michele Pagano*, Avram Hershko

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

436 Scopus citations

Abstract

The cyclin-dependent kinase (CDK) inhibitor p27 is degraded in late G1 phase by the ubiquitin pathway1, allowing CDK activity to drive cells into S phase2. Ubiquitinylation of p27 requires its phosphorylation at Thr 187 (refs 3, 4) and subsequent recognition by S-phase kinase associated protein 2 (Skp2; refs 5-8), a member of the F-box family of proteins that associates with Skp1, Cul-1 and ROC1/Rbx1 to form an SCF ubiquitin ligase complex9. However, in vitro ligation of p27 to ubiquitin could not be reconstituted by known purified components of the SCFSkp2 complex. Here we show that the missing factor is CDK subunit 1 (Cks1), which belongs to the highly conserved Suc1/Cks family of proteins that bind to some CDKs and phosphorylated proteins and are essential for cell-cycle progression. Human Cks1, but not other members of the family, reconstitutes ubiquitin ligation of p27 in a completely purified system, binds to Skp2 and greatly increases binding of T187-phosphorylated p27 to Skp2. Our results represent the first evidence that an SCF complex requires an accessory protein for activity as well as for binding to its phosphorylated substrate.

Original languageEnglish
Pages (from-to)321-324
Number of pages4
JournalNature Cell Biology
Volume3
Issue number3
DOIs
StatePublished - 2001
Externally publishedYes

Funding

FundersFunder number
Human Frontier Science Program Organization
Irma T. Hirschl Scholarship
National Cancer Institute of Canada
National Institutes of Health
International Union Against Cancer
Israel Science Foundation

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