title: Membrane-displayed peptide ligand activates the pheromone response pathway in Saccharomyces cerevisiae.
abstract: The budding yeast, Saccharomyces cerevisiae, is an attractive host for studying G protein-coupled receptors (GPCRs). We developed a system in which a peptide ligand specific for GPCR is displayed on yeast plasma membrane. The model system described here is based on yeast plasma membrane display of an analogue of α-factor, which is a peptide ligand for Ste2p, the GPCR that activates the yeast pheromone response pathway. α-Factor analogues, containing linkers of varying lengths and produced in yeast cells, became attached to the cell plasma membrane by linking to the glycosylphosphatidylinositol (GPI)-anchored plasma membrane protein Yps1p. We were able to demonstrate that an optimized α-factor analogue activated the pheromone response pathway in S. cerevisiae, as assessed by a fluorescent reporter assay. Furthermore, it was shown that linker length strongly influenced signalling pathway activation. To our knowledge, this is the first report documenting functional signalling by a plasma membrane-displayed ligand in S. cerevisiae.
authors.affiliation: Research Fellow of the Japan Society for the Promotion of Science, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Hara
authors.forename: Takuya
authors.initials: T
authors.lastname: Ono
authors.forename: Kouichi
authors.initials: K
authors.lastname: Kuroda
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Ueda
mesh_terms: D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D008024:Ligands; D010455:Peptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D010675:Pheromones / Q000494:pharmacology*; D043562:Receptors, G-Protein-Coupled / Q000037:antagonists & inhibitors / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D008024:Ligands; D010455:Peptides; D010675:Pheromones; D043562:Receptors, G-Protein-Coupled