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PMID: 14742714 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Identification of protein complexes required for efficient sister chromatid cohesion.

Molecular biology of the cell ·Vol. 15 ·No. 4 ·2004-04-00 ·Pages 1736-45

Mayer ML, Pot I, Chang M, Xu H, Aneliunas V, Kwok T, Newitt R, Aebersold R, Boone C, Brown GW, Hieter P

Abstract

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.

MeSH Terms
Chromatids/ultrastructure Chromosomes, Fungal Epitopes/chemistry Fungal Proteins/physiology Gene Deletion Genetic Techniques Genome, Fungal Green Fluorescent Proteins Luminescent Proteins/metabolism Mass Spectrometry Mutation Oligonucleotide Array Sequence Analysis Precipitin Tests Saccharomyces cerevisiae/physiology Saccharomyces cerevisiae Proteins/physiology
Chemicals
Epitopes Fungal Proteins Luminescent Proteins Saccharomyces cerevisiae Proteins Green Fluorescent Proteins
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Mayer Melanie L
Centre for Molecular Medicine and Therapeutics, University of British Columbia, Vancouver, British Columbia, Canada V5Z 4H4.
Pot Isabelle
Chang Michael
Xu Hong
Aneliunas Victoria
Kwok Teresa
Newitt Rick
Aebersold Ruedi
Boone Charles
Brown Grant W
Hieter Philip
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e03-08-0619
Published
2004-04-00
Epub
2004-00-23
Pages
1736-45
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC379271
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.e03-08-0619 PMC Full Text View journal details
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