Comprehensive genome-wide characterization of the pheromone response.
Corey Nislow (1), Angela M. Chu (1), Fred Naider (2), Ronald W. Davis (1), Guri Giaever (1)
(1) Department of Biochemistry, Stanford Genome Tech Center, 855 California Ave., Palo Alto, CA, 94304, United States;
(2) Department of Chemistry, College of Staten Island and Institute for Macromolecular Assemblies, City University of New York, Staten Island, New York 10314
Upon exposure to mating pheromone, cells of the opposite mating type undergo a series of morphological, biological and genetic changes. To gain additional understanding of how the cell coordinates the diverse physiological responses induced by mating pheromone, we have applied a parallel genome-wide approach. To accomplish this, we challenged a pooled collection of ~4800 bar-coded haploid deletion strains with either alpha-factor or a-factor and detected those mutants that were resistant to pheromone-induced growth arrest. We uncovered ~30 strains deleted for genes known to be involved in pheromone response, demonstrating that the parallel growth assay is robust. We also identified ~50 strains deleted for genes not previously known to be part of the pathway, and each of these novel strains were confirmed individually. Each strain was then subjected to a battery of phenotypic tests, including morphological and cytoskeletal characterization. We found that the responses to alpha-factor and a-factor were similar, but not identical. In a follow-up assay the deletion pools were challenged with a number of environmental stresses known to induce programmed cell death in Saccharomyces. A comparison of these datasets reveals a genome-wide perspective of those elements shared by both the mating response and cell death.
Return to YGM 2004 Home at SGD