- PhenoM: a database of morphological phenotypes caused by mutation of essential genes in Saccharomyces cerevisiae. [PMID: 22009677]
Ke Jin, Jingjing Li, Frederick S Vizeacoumar, Zhijian Li, Renqiang Min, Lee Zamparo, Franco J Vizeacoumar, Alessandro Datti, Brenda Andrews, Charles Boone, Zhaolei Zhang
Nucleic acids research 2012:40(Database issue)
6 Citations (Google Scholar as of 2016-01-14)
Abstract: About one-fifth of the genes in the budding yeast are essential for haploid viability and cannot be functionally assessed using standard genetic approaches such as gene deletion. To facilitate genetic analysis of essential genes, we and others have assembled collections of yeast strains expressing temperature-sensitive (ts) alleles of essential genes. To explore the phenotypes caused by essential gene mutation we used a panel of genetically engineered fluorescent markers to explore the morphology of cells in the ts strain collection using high-throughput microscopy. Here, we describe the design and implementation of an online database, PhenoM (Phenomics of yeast Mutants), for storing, retrieving, visualizing and data mining the quantitative single-cell measurements extracted from micrographs of the ts mutant cells. PhenoM allows users to rapidly search and retrieve raw images and their quantified morphological data for genes of interest. The database also provides several data-mining tools, including a PhenoBlast module for phenotypic comparison between mutant strains and a Gene Ontology module for functional enrichment analysis of gene sets showing similar morphological alterations. The current PhenoM version 1.0 contains 78,194 morphological images and 1,909,914 cells covering six subcellular compartments or structures for 775 ts alleles spanning 491 essential genes. PhenoM is freely available at http://phenom.ccbr.utoronto.ca/.
- Systematic exploration of essential yeast gene function with temperature-sensitive mutants. [PMID: 21441928]
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 Datti, Guri Giaever, Corey Nislow, Chris Bulawa, Chad L Myers, Michael Costanzo, Anne-Claude Gingras, Zhaolei Zhang, Anders Blomberg, Kerry Bloom, Brenda Andrews, Charles Boone
Nature biotechnology 2011:29(4)
128 Citations (Google Scholar as of 2016-01-18)
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 (â¼45%) of the 1,101 essential yeast genes, with â¼30% 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.
- Integrating high-throughput genetic interaction mapping and high-content screening to explore yeast spindle morphogenesis. [PMID: 20065090]
Franco J Vizeacoumar, Nydia van Dyk, Frederick S Vizeacoumar, Vincent Cheung, Jingjing Li, Yaroslav Sydorskyy, Nicolle Case, Zhijian Li, Alessandro Datti, Corey Nislow, Brian Raught, Zhaolei Zhang, Brendan Frey, Kerry Bloom, Charles Boone, Brenda J Andrews
The Journal of cell biology 2010:188(1)
62 Citations (Google Scholar as of 2016-01-18)
Abstract: We describe the application of a novel screening approach that combines automated yeast genetics, synthetic genetic array (SGA) analysis, and a high-content screening (HCS) system to examine mitotic spindle morphogenesis. We measured numerous spindle and cellular morphological parameters in thousands of single mutants and corresponding sensitized double mutants lacking genes known to be involved in spindle function. We focused on a subset of genes that appear to define a highly conserved mitotic spindle disassembly pathway, which is known to involve Ipl1p, the yeast aurora B kinase, as well as the cell cycle regulatory networks mitotic exit network (MEN) and fourteen early anaphase release (FEAR). We also dissected the function of the kinetochore protein Mcm21p, showing that sumoylation of Mcm21p regulates the enrichment of Ipl1p and other chromosomal passenger proteins to the spindle midzone to mediate spindle disassembly. Although we focused on spindle disassembly in a proof-of-principle study, our integrated HCS-SGA method can be applied to virtually any pathway, making it a powerful means for identifying specific cellular functions.