Publication

Identification of protein complexes required for efficient sister chromatid cohesion

Mayer, ML., Pot, A., Chang, M., Xu, H., Aneliunas, V., Kwok, T., Newitt, R., Aebersold, R., Boone, C., Brown, GW. & Hieter, P., Apr-2004, In : Molecular Biology of the Cell. 15, 4, p. 1736-1745 10 p.

Research output: Contribution to journalArticleAcademicpeer-review

  • ML Mayer
  • A. Pot
  • M Chang
  • H Xu
  • Victoria Aneliunas
  • T Kwok
  • R Newitt
  • R Aebersold
  • C Boone
  • GW Brown
  • P Hieter

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.

Original languageEnglish
Pages (from-to)1736-1745
Number of pages10
JournalMolecular Biology of the Cell
Volume15
Issue number4
Publication statusPublished - Apr-2004
Externally publishedYes

    Keywords

  • Chromatids, Chromosomes, Fungal, Epitopes, Fungal Proteins, Gene Deletion, Genetic Techniques, Genome, Fungal, Green Fluorescent Proteins, Luminescent Proteins, Mass Spectrometry, Mutation, Oligonucleotide Array Sequence Analysis, Precipitin Tests, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins

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