Identification of a glycoprotein involved in cell cycle progression in yeast.
The molecular events of start, the regulatory step that commits yeast cells to DNA replication, have recently begun to be investigated. One of the gene products required for completion of start has been found to have a significant structural homology with oncogenes endowed with protein kinase activity. Our experiments provide data on the biosynthetic pathway of a previously identified labile protein (p100, molecular weight 100,000, isoelectric point of approximately 4.8-5) involved in cell cycle progression at start, which appears to be specifically made during the release from cell cycle arrest of a temperature-sensitive mutant (cdc25) of Saccharomyces cerevisiae. On two-dimensional gel, p100 migrates very close to another 100-kDa labile protein (p100*) which behaves as a cell cycle modulated protein with reduced synthesis in G1. Pulse and chase labeling of protein with [35S]methionine suggests that both p100 and p100* are processed to a protein (p115) of slightly higher molecular weight (Mr = 115,000). Peptide mapping analysis indicates that p100 and p100 yield identical maps and that both p100 and p100* are very much similar to p115. p115 is a glycosylated protein as shown by a labeling experiment with [3H]glucosamine and by the fact that the synthesis of both p100 and p115 is inhibited if cells are cultured in the presence of tunicamycin. A protein having the same heterogeneous aspect of migration on sodium dodecyl sulfate-polyacrylamide gel and the same apparent molecular weight and isoelectric point of p115 is abundantly present in a preparation of membranes from S. cerevisiae and the isolated radioactive p115 comigrates with it. Taken together these results favor the idea that terminal glycosylation of both p100 and p100* gives rise to the fully glycosylated p115 protein which appears to be a membrane-associated protein.
D002453:Cell Cycle; D003513:Cycloheximide / Q000494:pharmacology; D005656:Fungal Proteins / Q000032:analysis / Q000096:biosynthesis*; D006023:Glycoproteins / Q000032:analysis / Q000096:biosynthesis*; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013462:Sulfur Radioisotopes; D014415:Tunicamycin / Q000494:pharmacology
D005656:Fungal Proteins; D006023:Glycoproteins; D013462:Sulfur Radioisotopes; D014415:Tunicamycin; D003513:Cycloheximide; D008715:Methionine