Effects of expression of mammalian G alpha and hybrid mammalian-yeast G alpha proteins on the yeast pheromone response signal transduction pathway.
Scg1, the product of the Saccharomyces cerevisiae SCG1 (also called GPA1) gene, is homologous to the alpha subunits of G proteins involved in signal transduction in mammalian cells. Scg1 negatively controls the pheromone response pathway in haploid cells. Either pheromonal activation or an scg1 null mutation relieves the negative control and leads to an arrest of cell growth in the G1 phase of the cell cycle. Expression of rat G alpha s was previously shown to complement the growth defect of scg1 null mutants while not allowing mating. We have extended this analysis to examine the effects of the short form of G alpha s (which lacks 15 amino acids present in the long form), G alpha i2, G alpha o, and Scg1-mammalian G alpha hybrids. In addition, we have found that constructs able to complement scg1 are also able to inhibit the response to pheromone and mating when expressed in a wild-type SCG1 strain. Overexpression of Scg1 has a similar inhibitory effect. These results are consistent with a model proposed for the action of Scg1 as the alpha component of a heterotrimeric G protein in which the beta gamma component (Ste4/Ste18) activates the pheromone response after dissociation from Scg1. They suggest that the G alpha constructs able to complement scg1 can interact with beta gamma to prevent activation of the pathway but are unable to interact with pheromone receptors to activate the pathway.
Kang
Y S
YS
Department of Molecular Genetics and Microbiology, University of Massachusetts Medical Center, Worcester 01655.
Kane
J
J
Kurjan
J
J
Stadel
J M
JM
Tipper
D J
DJ
D000818:Animals; D001483:Base Sequence; D003433:Crosses, Genetic; D019204:GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D046911:Macromolecular Substances; D000072235:Mating Factor; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D010455:Peptides / Q000502:physiology*; D010675:Pheromones / Q000502:physiology*; D010957:Plasmids; D055503:Protein Multimerization; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000502:physiology; D015398:Signal Transduction
D046911:Macromolecular Substances; D015345:Oligonucleotide Probes; D010455:Peptides; D010675:Pheromones; D011993:Recombinant Fusion Proteins; D000072235:Mating Factor; D019204:GTP-Binding Proteins