Genetic screens in yeast to identify mammalian nonreceptor modulators of G-protein signaling.
We describe genetic screens in Saccharomyces cerevisiae designed to identify mammalian nonreceptor modulators of G-protein signaling pathways. Strains lacking a pheromone-responsive G-protein coupled receptor and expressing a mammalian-yeast Galpha hybrid protein were made conditional for growth upon either pheromone pathway activation (activator screen) or pheromone pathway inactivation (inhibitor screen). Mammalian cDNAs that conferred plasmid-dependent growth under restrictive conditions were identified. One of the cDNAs identified from the activator screen, a human Ras-related G protein that we term AGS1 (for activator of G-protein signaling), appears to function by facilitating guanosine triphosphate (GTP) exchange on the heterotrimeric Galpha. A cDNA product identified from the inhibitor screen encodes a previously identified regulator of G-protein signaling, human RGS5.
Cismowski
M J
MJ
Cadus Pharmaceutical Corporation, 777 Old Saw Mill River Road, Tarrytown 10591, NY, USA.
Takesono
A
A
Ma
C
C
Lizano
J S
JS
Xie
X
X
Fuernkranz
H
H
Lanier
S M
SM
Duzic
E
E
D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics; D004843:Epistasis, Genetic; D019204:GTP-Binding Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D015723:Gene Library; D005818:Genetic Engineering; D006801:Humans; D010641:Phenotype; D010675:Pheromones / Q000378:metabolism; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics*; D012641:Selection, Genetic; D015398:Signal Transduction / Q000235:genetics*; D014170:Transformation, Genetic; D018631:ras Proteins / Q000235:genetics / Q000302:isolation & purification*
D018076:DNA, Complementary; D010675:Pheromones; C400132:RASD1 protein, human; D019204:GTP-Binding Proteins; D018631:ras Proteins