Systematic exploration of essential yeast gene function with temperature-sensitive mutants

Zhijian Li, Franco J. Vizeacoumar, Sondra Bahr, Jingjing Li, Jonas Warringer, Frederick S. Vizeacoumar, Renqiang Min, Benjamin Vandersluis, Jeremy Bellay, Michael Devit, James A. Fleming, Andrew Stephens, Julian Haase, Zhen Yuan Lin, Anastasia Baryshnikova, Hong Lu, Zhun Yan, Ke Jin, Sarah Barker, Alessandro DattiGuri Giaever, Corey Nislow, Chris Bulawa, Chad L. Myers, Michael Costanzo, Anne Claude Gingras, Zhaolei Zhang, Anders Blomberg, Kerry Bloom, Brenda Andrews, Charles Boone

Research output: Contribution to journalArticlepeer-review

221 Scopus citations

Abstract

Conditional temperature-sensitive (ts) mutations are valuable reagents for studying essential genes in the yeast Saccharomyces cerevisiae. We constructed 787 ts strains, covering 497 (∼45%) of the 1,101 essential yeast genes, with ∼30% of the genes represented by multiple alleles. All of the alleles are integrated into their native genomic locus in the S288C common reference strain and are linked to a kanMX selectable marker, allowing further genetic manipulation by synthetic genetic array (SGA)-based, high-throughput methods. We show two such manipulations: barcoding of 440 strains, which enables chemical-genetic suppression analysis, and the construction of arrays of strains carrying different fluorescent markers of subcellular structure, which enables quantitative analysis of phenotypes using high-content screening. Quantitative analysis of a GFP-tubulin marker identified roles for cohesin and condensin genes in spindle disassembly. This mutant collection should facilitate a wide range of systematic studies aimed at understanding the functions of essential genes.

Original languageEnglish (US)
Pages (from-to)361-367
Number of pages7
JournalNature biotechnology
Volume29
Issue number4
DOIs
StatePublished - Apr 2011

Bibliographical note

Funding Information:
We thank the members of the Boone, Andrews and Bloom laboratory for their input and discussions. We thank S. Biggins (Fred Hutchinson Cancer Research Center) for her insights on cohesin and condensin components. We thank M. Zackrisson (University of Gothenburg) for statistical support. The Sli15-6A-GFP construct was a gift from E. Schiebel (University of Heidelberg) and the Mad1-NLS construct was a gift from R. Wozniak (University of Alberta). We also thank Jennifer Reginold for manual inspection of HCS images. F.J.V. was supported by a postdoctoral fellowship from the Best Foundation. B.A. and C.B. were supported by Genome Canada through the Ontario Genomics Institute as per research agreement 2004-OGI-3-01 and the Canadian Institutes of Health Research (MOP-97939).

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