Cyclic AMP responses are suppressed in mammalian cells expressing the yeast low Km cAMP-phosphodiesterase gene

J Biol Chem. 1990 Apr 5;265(10):5840-6.

Abstract

A genomic DNA fragment from Saccharomyces cerevisiae which contains the SRA5 (=PDE2) gene, coding for a low Km cAMP-phosphodiesterase, was transfected into Chinese hamster ovary cells. Clones carring the cAMP-phosphodiesterase gene were capable of growth in the presence of cholera toxin, which slows the growth of untransfected cells by elevating their cAMP levels. The cholera toxin-resistant transfected cell lines expressed high levels of cAMP-phosphodiesterase mRNA and cAMP-phosphodiesterase activity. Basal intracellular cAMP levels were not significantly affected by the presence of the yeast cAMP-phosphodiesterase gene, but elevation of cAMP levels in response to cholera toxin or prostaglandin E1 was suppressed. Induction of the cAMP-responsive tyrosine aminotransferase promoter by cholera toxin was also blocked in cell lines carrying the yeast cAMP-phosphodiesterase gene. Cholera toxin-resistant transfected cell lines were sensitive to the growth inhibitory effects of N6,02'-dibutyryladenosine 3',5'-monophosphate, which can be used to bypass the effects of the yeast cAMP-phosphodiesterase.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 1-Methyl-3-isobutylxanthine / pharmacology
  • 3',5'-Cyclic-AMP Phosphodiesterases / genetics*
  • Alprostadil / pharmacology
  • Animals
  • Bucladesine / pharmacology
  • Cell Division / drug effects
  • Cell Line
  • Cholera Toxin / pharmacology
  • Cricetinae
  • Cyclic AMP / metabolism
  • Cyclic AMP / pharmacology*
  • Gene Expression*
  • Genes, Fungal*
  • Kinetics
  • Promoter Regions, Genetic / genetics
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics*
  • Signal Transduction
  • Transcription, Genetic
  • Transfection
  • Transformation, Genetic

Substances

  • Bucladesine
  • Cholera Toxin
  • Cyclic AMP
  • 3',5'-Cyclic-AMP Phosphodiesterases
  • Alprostadil
  • 1-Methyl-3-isobutylxanthine