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Molecular and Cellular Biology, October 2004, p. 8938-8950, Vol. 24, No. 20
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.20.8938-8950.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

The Interaction between Sgt1p and Skp1p Is Regulated by HSP90 Chaperones and Is Required for Proper CBF3 Assembly{dagger}

Linda B. Lingelbach and Kenneth B. Kaplan*

Section of Molecular and Cellular Biology, University of California Davis, Davis, California

Received 24 February 2004/ Returned for modification 30 March 2004/ Accepted 26 July 2004

Sgt1p is a well-conserved protein proposed to be involved in a number of cellular processes. Genetic studies of budding yeast suggest a role for SGT1 in signal transduction, cell cycle advance, and chromosome segregation. Recent evidence has linked Sgt1p to HSP90 chaperones, although the precise relationship between these proteins is unclear. To further explore the role of Sgt1p in these processes, we have characterized the interactions among Sgt1p, the inner kinetochore complex CBF3, and HSP90 chaperones. We show that the amino terminus of Sgt1p interacts with CBF3 subunits Skp1p and Ctf13p. HSP90 interacts with Sgt1p and, in combination with the carboxy terminus of Sgt1p, regulates the interaction between Sgt1p and Skp1p in a nucleotide-dependent manner. While the Sgt1p-Skp1p interaction is required for CBF3 assembly, mutations that stabilize this interaction prevent the turnover of protein complexes important for CBF3 assembly. We propose that HSP90 and Sgt1p act together as a molecular switch, maintaining transient interactions required to balance protein complex assembly with turnover.

* Corresponding author. Mailing address: Section of Molecular and Cellular Biology, UC Davis, Davis, CA 95616. Phone: (530) 754-5044. Fax: (530) 752-3085. E-mail: kbkaplan{at}ucdavis.edu.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, October 2004, p. 8938-8950, Vol. 24, No. 20
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.20.8938-8950.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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