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

Michiel M. Van Lockeren Campagne, Erica Wu, Robert D. Fleischmann, Michael M. Gottesman, Kevin W. Chason, Richard H. Kessin

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

A genomic DNA fragment from Saccharomyces cereuisiae which contains the SRA5 (=PDE2) gene, coding for a low Km cAMP-phosphodiesterase, was transfected into Chinese hamster ovary cells. Clones carrying 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 El was suppressed. Induction of the c AMP-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,O2′-dibutyryladenosine 3′,5′-monophosphate, which can be used to bypass the effects of the yeast cAMP-phosphodiesterase.

Original languageEnglish
Pages (from-to)5840-5846
Number of pages7
JournalJournal of Biological Chemistry
Volume265
Issue number10
StatePublished - 1990
Externally publishedYes

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