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Vol. 15, Issue 4, 1736-1745, April 2004
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¶ #
* Centre for Molecular Medicine and Therapeutics, University of British Columbia, Vancouver, British Columbia, Canada V5Z 4H4;
¶ Biotechnology Laboratory, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3;
Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3;
Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada M5S 1A8;
Department of Medical Genetics and Microbiology, University of Toronto, Toronto, Ontario, Canada M5S 1A8 and Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, Canada M5G 1L6; and
|| Institute for Systems Biology, Seattle, Washington 98105
Submitted August 22, 2003;
Revised November 9, 2003;
Accepted November 19, 2003
Monitoring Editor: Frank Solomon
Ctf8p is a component of Ctf18-RFC, an alternative replication factor C-like complex required for efficient sister chromatid cohesion in Saccharomyces cerevisiae. We performed synthetic genetic array (SGA) analysis with a ctf8 deletion strain as a primary screen to identify other nonessential genes required for efficient sister chromatid cohesion. We then assessed proficiency of cohesion at three chromosomal loci in strains containing deletions of the genes identified in the ctf8 SGA screen. Deletion of seven genes (CHL1, CSM3, BIM1, KAR3, TOF1, CTF4, and VIK1) resulted in defective sister chromatid cohesion. Mass spectrometric analysis of immunoprecipitated complexes identified a physical association between Kar3p and Vik1p and an interaction between Csm3p and Tof1p that we confirmed by coimmunoprecipitation from cell extracts. These data indicate that synthetic genetic array analysis coupled with specific secondary screens can effectively identify protein complexes functionally related to a reference gene. Furthermore, we find that genes involved in mitotic spindle integrity and positioning have a previously unrecognized role in sister chromatid cohesion.
# Corresponding author. E-mail address: hieter{at}cmmt.ubc.ca.
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