title: Chimeric yeast G-protein α subunit harboring a 37-residue C-terminal gustducin-specific sequence is functional in Saccharomyces cerevisiae.
abstract: Despite many recent studies of G-protein-coupled receptor (GPCR) structures, it is not yet well understood how these receptors activate G proteins. The GPCR assay using baker's yeast, Saccharomyces cerevisiae, is an effective experimental model for the characterization of GPCR-Gα interactions. Here, using the yeast endogenous Gα protein (Gpa1p) as template, we constructed various chimeric Gα proteins with a region that is considered to be necessary for interaction with mammalian receptors. The signaling assay using the yeast pheromone receptor revealed that the chimeric Gα protein harboring 37 gustducin-specific amino acid residues at its C-terminus (GPA1/gust37) maintained functionality in yeast. In contrast, GPA1/gust44, a variant routinely used in mammalian experimental systems, was not functional.
authors.affiliation: Japan Society for the Promotion of Science, Sakyo-ku, Kyoto, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Hara
authors.forename: Yuko
authors.initials: Y
authors.lastname: Inada
authors.forename: Takuya
authors.initials: T
authors.lastname: Ono
authors.forename: Kouichi
authors.initials: K
authors.lastname: Kuroda
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Yasuda-Kamatani
authors.forename: Masaji
authors.initials: M
authors.lastname: Ishiguro
authors.forename: Takaharu
authors.initials: T
authors.lastname: Tanaka
authors.forename: Takumi
authors.initials: T
authors.lastname: Misaka
authors.forename: Keiko
authors.initials: K
authors.lastname: Abe
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Ueda
mesh_terms: D000595:Amino Acid Sequence; D044385:GTP-Binding Protein alpha Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015293:Transducin / Q000235:genetics*
chemical_list: D044385:GTP-Binding Protein alpha Subunits; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C075192:gustducin; D015293:Transducin