Reconstitution of G1 cyclin ubiquitination with complexes containing SCF(Grr1) and Rbx1

Dorota Skowyra, Deanna M. Koepp, Takumi Kamura, Michael N. Conrad, Ronald C. Conaway, Joan Weliky Conaway, Stephen J. Elledge, J. Wade Harper

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345 Scopus citations

Abstract

Control of cyclin levels is critical for proper cell cycle regulation. In yeast, the stability of the G1 cyclin Cln1 is controlled by phosphorylation-dependent ubiquitination. Here it is shown that this reaction can be reconstituted in vitro with an SCF E3 ubiquitin ligase complex. Phosphorylated Cln1 was ubiquitinated by SCF (Skp1-Cdc53-F-box protein) complexes containing the F-box protein Grr1, Rbx1, and the E2 Cdc34. Rbx1 promotes association of Cdc34 with Cdc53 and stimulates Cdc34 auto- ubiquitination in the context of Cdc53 or SCF complexes. Rbx1, which is also a component of the von Hippel-Lindau tumor suppressor complex, may define a previously unrecognized class of E3-associated proteins.

Original languageEnglish (US)
Pages (from-to)662-665
Number of pages4
JournalScience
Volume284
Issue number5414
DOIs
StatePublished - Apr 23 1999

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Skowyra, D., Koepp, D. M., Kamura, T., Conrad, M. N., Conaway, R. C., Conaway, J. W., Elledge, S. J., & Harper, J. W. (1999). Reconstitution of G1 cyclin ubiquitination with complexes containing SCF(Grr1) and Rbx1. Science, 284(5414), 662-665. https://doi.org/10.1126/science.284.5414.662