Phosphorylation of RAS1 and RAS2 proteins in Saccharomyces cerevisiae.
RAS1 and RAS2 proteins of Saccharomyces cerevisiae are guanine nucleotide-binding proteins involved in the regulation of adenylate cyclase. In this paper, we report that these proteins are phosphorylated. The phosphorylation of RAS1 protein is demonstrated by treating with alkaline phosphatase as well as by labeling with [32P]orthophosphate. The phosphorylation occurs exclusively on serine residues and phosphorylated RAS1 protein is predominantly membrane localized. The phosphorylation of RAS2 protein is demonstrated by similar 32P-labeling experiments. The phosphorylation occurs exclusively on serine residues and phosphopeptide analyses suggest that only two major phosphorylated tryptic peptides are generated from the RAS2 protein. These results provide evidence for the phosphorylation of RAS proteins in vivo. Furthermore, our demonstration that the phosphorylation occurs exclusively on serine residues and that the RAS2 protein contains only two major phosphorylated tryptic peptides argues that the phosphorylation may be physiologically significant.
Cobitz
A R
AR
Department of Biochemistry and Molecular Biology, University of Chicago, IL 60637.
Yim
E H
EH
Brown
W R
WR
Perou
C M
CM
Tamanoi
F
F
D000469:Alkaline Phosphatase; D000596:Amino Acids / Q000032:analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010748:Phosphopeptides / Q000032:analysis; D010766:Phosphorylation; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D014357:Trypsin; D018631:ras Proteins
D000596:Amino Acids; D005656:Fungal Proteins; D010446:Peptide Fragments; D010748:Phosphopeptides; D029701:Saccharomyces cerevisiae Proteins; D000469:Alkaline Phosphatase; D014357:Trypsin; D019204:GTP-Binding Proteins; C489014:RAS1 protein, S cerevisiae; C489015:RAS2 protein, S cerevisiae; D018631:ras Proteins