Overexpression of the CDC25 gene, an upstream element of the RAS/adenylyl cyclase pathway in Saccharomyces cerevisiae, allows immunological identification and characterization of its gene product

Biochem Biophys Res Commun. 1990 Oct 15;172(1):61-9. doi: 10.1016/s0006-291x(05)80173-1.

Abstract

The product of the START gene CDC25, an upstream element of the RAS/adenylyl cyclase pathway in Saccharomyces cerevisiae, was identified using specific antibodies raised against a chimeric beta-galactosidase/CDC25 protein. The CDC25 protein is poorly expressed and can be detected only when the CDC25 gene is overexpressed under the control of the galactose-inducible GAL1-10 strong promoter elements. It has a molecular weight of 180,000, is not glycosylated and is strongly associated with the particulate fraction. After deletion of residues 1255-1550 the protein is found in the soluble fraction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases / genetics*
  • Antibodies
  • Cell Cycle Proteins*
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Fungal Proteins / analysis
  • Fungal Proteins / genetics*
  • Fungal Proteins / immunology
  • Gene Expression Regulation, Fungal*
  • Genes, Fungal*
  • Immunoblotting
  • Molecular Weight
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / immunology
  • Restriction Mapping
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins
  • beta-Galactosidase / analysis
  • beta-Galactosidase / genetics
  • beta-Galactosidase / immunology
  • ras Proteins*
  • ras-GRF1*

Substances

  • Antibodies
  • CDC25 protein, S cerevisiae
  • Cell Cycle Proteins
  • Fungal Proteins
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • ras-GRF1
  • beta-Galactosidase
  • ras Proteins
  • Adenylyl Cyclases