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Home/Research/Paper Chase/Recombinant fragment of protein kinase inhibitor blocks cyclic AMP-dependent gene transcription.
Recombinant fragment of protein kinase inhibitor blocks cyclic AMP-dependent gene transcription.
Science.Oct 23, 1987;238(4826):530-3.
Grove JR, Price DJ, Goodman HM, Avruch J.
Howard Hughes Medical Institute, Harvard Medical School, Boston, MA 02114.
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.