title: Integrating high-throughput genetic interaction mapping and high-content screening to explore yeast spindle morphogenesis.
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.
authors.affiliation: Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario M5S 3E1, Canada.
authors.forename: Franco J
authors.initials: FJ
authors.lastname: Vizeacoumar
authors.forename: Nydia
authors.initials: N
authors.lastname: van Dyk
authors.forename: Frederick
authors.initials: F
authors.lastname: S Vizeacoumar
authors.forename: Vincent
authors.initials: V
authors.lastname: Cheung
authors.forename: Jingjing
authors.initials: J
authors.lastname: Li
authors.forename: Yaroslav
authors.initials: Y
authors.lastname: Sydorskyy
authors.forename: Nicolle
authors.initials: N
authors.lastname: Case
authors.forename: Zhijian
authors.initials: Z
authors.lastname: Li
authors.forename: Alessandro
authors.initials: A
authors.lastname: Datti
authors.forename: Corey
authors.initials: C
authors.lastname: Nislow
authors.forename: Brian
authors.initials: B
authors.lastname: Raught
authors.forename: Zhaolei
authors.initials: Z
authors.lastname: Zhang
authors.forename: Brendan
authors.initials: B
authors.lastname: Frey
authors.forename: Kerry
authors.initials: K
authors.lastname: Bloom
authors.forename: Charles
authors.initials: C
authors.lastname: Boone
authors.forename: Brenda J
authors.initials: BJ
authors.lastname: Andrews
mesh_terms: D057205:Automation, Laboratory; D005821:Genetic Techniques; D009154:Mutation; D011485:Protein Binding; D025941:Protein Interaction Mapping; D025842:SUMO-1 Protein / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000166:cytology* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D008941:Spindle Apparatus / Q000737:chemistry* / Q000235:genetics
chemical_list: D025842:SUMO-1 Protein; D029701:Saccharomyces cerevisiae Proteins