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

Enzo Martegani, Marco Vanoni, Renata Zippel, Paola Coccetti, Riccardo Brambilla, Cristina Ferrari, Emmapaola Sturani, Lilia Alberghina

Research output: Contribution to journalArticlepeer-review

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 CDCZS-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 CDC25 , 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 ∼ 1700 and 5200 nucleotides. Transcripts were found also in mouse heart and at a lower level in liver and spleen.

Original languageEnglish
Pages (from-to)2151-2157
Number of pages7
JournalEMBO Journal
Volume11
Issue number6
Publication statusPublished - 1992

Keywords

  • CDC25 complementation
  • Mouse GNRP
  • RAS activator

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

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