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

doi:10.1126/science.284.5414.662

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

Genetics, Cell Biology and Development (CBS)

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359 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 language	English (US)
Pages (from-to)	662-665
Number of pages	4
Journal	Science
Volume	284
Issue number	5414
DOIs	https://doi.org/10.1126/science.284.5414.662
State	Published - Apr 23 1999

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title = "Reconstitution of G1 cyclin ubiquitination with complexes containing SCF(Grr1) and Rbx1",

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.",

author = "Dorota Skowyra and Koepp, {Deanna M.} and Takumi Kamura and Conrad, {Michael N.} and Conaway, {Ronald C.} and Conaway, {Joan Weliky} and Elledge, {Stephen J.} and Harper, {J. Wade}",

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T1 - Reconstitution of G1 cyclin ubiquitination with complexes containing SCF(Grr1) and Rbx1

AU - Skowyra, Dorota

AU - Koepp, Deanna M.

AU - Kamura, Takumi

AU - Conrad, Michael N.

AU - Conaway, Ronald C.

AU - Conaway, Joan Weliky

AU - Elledge, Stephen J.

AU - Harper, J. Wade

PY - 1999/4/23

Y1 - 1999/4/23

N2 - 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.

AB - 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.

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EP - 665

JO - Science

JF - Science

IS - 5414

ER -

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

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