title: Composite synthetic lethal identification of membrane traffic inhibitors.
abstract: Small molecule inhibitors provide powerful tools to characterize highly dynamic and complex eukaryotic cell pathways such as those mediating membrane traffic. However, a lack of easy and generalizable assays has constrained identification of novel inhibitors despite availability of diverse chemical libraries. Here, we report a facile growth-based strategy in yeast to screen for pathway-specific inhibitors. The approach uses well characterized synthetic genetic growth defects to guide design of cells genetically sensitized for inhibition of chosen pathways. With this strategy, we identified a family of piperazinyl phenylethanone compounds as inhibitors of traffic between the trans-Golgi network (TGN) and endosomes that depends on the clathrin adaptor complex AP-1. The compounds did not significantly alter other trafficking pathways involving the TGN or endosomes, indicating specificity. Compound treatment also altered localization of AP-1 in mammalian cells. These previously uncharacterized inhibitors will be useful for future studies of clathrin-mediated transport in yeast, and potentially in other organisms. Furthermore, the easily automated technology should be adaptable for identification of inhibitors of other cellular processes.
authors.affiliation: Department of Biological Chemistry, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
authors.forename: Mara C
authors.initials: MC
authors.lastname: Duncan
authors.forename: David G
authors.initials: DG
authors.lastname: Ho
authors.forename: Jing
authors.initials: J
authors.lastname: Huang
authors.forename: Michael E
authors.initials: ME
authors.lastname: Jung
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Payne
mesh_terms: D033961:Adaptor Protein Complex 1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics*; D002470:Cell Survival; D002686:Chitin / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism*; D005804:Genes, Lethal / Q000235:genetics; D015394:Molecular Structure; D009154:Mutation / Q000235:genetics; D009930:Organic Chemicals / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D015003:Yeasts; D021601:trans-Golgi Network / Q000378:metabolism*
chemical_list: D033961:Adaptor Protein Complex 1; D033942:Adaptor Proteins, Vesicular Transport; C497361:Gga2 protein, S cerevisiae; D009930:Organic Chemicals; D029701:Saccharomyces cerevisiae Proteins; D002686:Chitin