Abstract
Transcriptional regulation by cyclic adenosine monophosphate (cAMP) in mammalian cells could be mediated by a phosphoprotein substrate of the cAMP-dependent protein kinase or, as in prokaryotes, by a cAMP-binding protein. Two synthetic genes that code for an active fragment of the protein inhibitor of this kinase and a mutant inactive fragment were constructed and used to distinguish these alternatives. Transient expression of the active peptide product specifically inhibited the cAMP-stimulated expression of a cotransfected reporter gene by more than 90 percent, whereas the expression of the inactive peptide did not alter cAMP-stimulated gene expression. The results indicate that an active kinase catalytic subunit is a necessary intermediate in the cAMP stimulation of gene transcription.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acetyltransferases / genetics
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Carrier Proteins / pharmacology*
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Chloramphenicol O-Acetyltransferase
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Cyanogen Bromide
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Cyclic AMP / pharmacology*
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DNA, Recombinant
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Escherichia coli / genetics
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Intracellular Signaling Peptides and Proteins*
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Nucleic Acid Hybridization
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Peptide Fragments / pharmacology*
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Phosphorylation
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Plasmids
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Protein Kinase Inhibitors
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Protein Kinases / metabolism
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RNA, Messenger / genetics
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Recombinant Proteins / pharmacology*
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Transcription, Genetic / drug effects*
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Transfection
Substances
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Carrier Proteins
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DNA, Recombinant
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Intracellular Signaling Peptides and Proteins
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Peptide Fragments
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Protein Kinase Inhibitors
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RNA, Messenger
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Recombinant Proteins
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protein kinase modulator
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Cyclic AMP
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Acetyltransferases
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Chloramphenicol O-Acetyltransferase
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Protein Kinases
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Cyanogen Bromide