Cloning by functional complementation of a mouse cDNA encoding a homologue of CDC25, a Saccharomyces cerevisiae RAS activator

EMBO J. 1992 Jun;11(6):2151-7. doi: 10.1002/j.1460-2075.1992.tb05274.x.

Abstract

In the yeast Saccharomyces cerevisiae genetic and biochemical evidence indicates that the product of the CDC25 gene activates the RAS/adenylyl cyclase/protein kinase A pathway by acting as a guanine nucleotide protein. Here we report the isolation of a mouse brain cDNA homologous to CDC25. The mouse cDNA, called CDC25Mm, complements specifically point mutations and deletion/disruptions of the CDC25 gene. In addition, it restores the cAMP levels and CDC25-dependent glucose-induced cAMP signalling in a yeast strain bearing a disruption of the CDC25 gene. The CDC25Mm-encoded protein is 34% identical with the catalytic carboxy terminal part of the CDC25 protein and shares significant homology with other proteins belonging to the same family. The protein encoded by CDC25Mm, prepared as a glutathione S-transferase fusion in Escherichia coli cells, activates adenylyl cyclase in yeast membranes in a RAS2-dependent manner. Northern blot analysis of mouse brain poly(A)+ RNA reveals two major transcripts of approximately 1700 and 5200 nucleotides. Transcripts were found also in mouse heart and at a lower level in liver and spleen.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases / metabolism*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Northern
  • Cell Cycle Proteins*
  • Cloning, Molecular
  • Cyclic AMP / metabolism
  • DNA / genetics*
  • DNA, Fungal / genetics
  • Enzyme Activation
  • Escherichia coli / genetics
  • Fungal Proteins / genetics*
  • Fungal Proteins / isolation & purification
  • Fungal Proteins / metabolism
  • GTP-Binding Proteins / genetics
  • Gene Expression Regulation, Fungal*
  • Genetic Complementation Test
  • Genotype
  • Glucose / pharmacology
  • Glutathione Transferase / genetics
  • Glutathione Transferase / isolation & purification
  • Glutathione Transferase / metabolism
  • Mice
  • Molecular Sequence Data
  • Plasmids
  • Poly A / genetics
  • Poly A / isolation & purification
  • RNA / genetics
  • RNA / isolation & purification
  • RNA, Messenger
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins*
  • Sequence Homology, Nucleic Acid
  • ras Proteins*
  • ras-GRF1*

Substances

  • CDC25 protein, S cerevisiae
  • Cell Cycle Proteins
  • DNA, Fungal
  • Fungal Proteins
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • ras-GRF1
  • Poly A
  • RNA
  • DNA
  • Cyclic AMP
  • Glutathione Transferase
  • GTP-Binding Proteins
  • RAS2 protein, S cerevisiae
  • ras Proteins
  • Adenylyl Cyclases
  • Glucose

Associated data

  • GENBANK/X59868
  • GENBANK/X64555
  • GENBANK/X64556
  • GENBANK/X64602
  • GENBANK/X64629
  • GENBANK/X64700
  • GENBANK/X67723
  • GENBANK/X67724
  • GENBANK/X67725
  • GENBANK/X67726