Antibody mimicking the action of RAS proteins on yeast adenylyl cyclase: implication for RAS-effector interaction.
Polyclonal antisera were raised against various subregions of Saccharomyces cerevisiae adenylyl cyclase in order to examine the molecular mechanism of interaction between adenylyl cyclase and RAS proteins. One of the antisera was found to activate adenylyl cyclase to an extent comparable to that activated by saturating amounts of yeast RAS2 protein produced in Escherichia coli. The stimulatory effect of this antiserum was shown to be additive with RAS2 protein when both antisera and RAS2 protein were present at low concentrations. At saturating amounts of RAS2 protein, the antisera did not exhibit additional stimulatory effects, suggesting that the actions of RAS2 protein and the antisera are complementary with each other. The antigenic determinant for the antibody involved in the activation was mapped to a 14-amino-acid segment, 1452-NSVDNGADVANLSY-1465, located between the leucine-rich repeats and the catalytic domain of adenylyl cyclase. Certain missense mutations affecting this 14-amino acid segment significantly reduced the response of adenylyl cyclase to both activating antibody and RAS proteins. These results suggest that this segment of adenylyl cyclase is intimately involved in the mechanism by which RAS proteins activate this downstream effector.
Suzuki
N
N
Department of Physiology, Kobe University School of Medicine, Japan.
Tsujino
K
K
Minato
T
T
Nishida
Y
Y
Okada
T
T
Kataoka
T
T
D000262:Adenylyl Cyclases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology / Q000378:metabolism*; D004789:Enzyme Activation; D000939:Epitopes; D005656:Fungal Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins; D015398:Signal Transduction; D018631:ras Proteins
D000906:Antibodies; D000939:Epitopes; D005656:Fungal Proteins; D029701:Saccharomyces cerevisiae Proteins; C489015:RAS2 protein, S cerevisiae; D018631:ras Proteins; D000262:Adenylyl Cyclases