Recombinant Fragment of Protein Kinase Inhibitor Blocks Cyclic AMP--Dependent Gene Transcription

• Published in: Science (American Association for the Advancement of Science) Vol. 238; no. 4826; pp. 530 - 533
• Main Authors: Grove, J. Russell, Price, Daniel J., Goodman, Howard M., Avruch, Joseph
• Format: Journal Article
• Language: English
• Published: Washington, DC The American Association for the Advancement of Science 23.10.1987; American Association for the Advancement of Science
• Subjects: Acetyltransferases - genetics; Amino acids; Biochemistry; Biological and medical sciences; Carrier Proteins - pharmacology; Cell extracts; Chloramphenicol O-Acetyltransferase; COS cells; Cyanogen Bromide; Cyclic AMP - pharmacology; DNA; DNA, Recombinant; Enzyme inhibitors; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Genes; Genetic engineering; Genetic transcription; Intracellular Signaling Peptides and Proteins; Legends; Medical research; Messenger RNA; Molecular and cellular biology; Molecular genetics; Nucleic Acid Hybridization; Peptide Fragments - pharmacology; Phosphoproteins; Phosphorylation; Plasmids; Protein Kinase Inhibitors; Protein kinases; Protein Kinases - metabolism; Protein research; Proteins; Recombinant Proteins - pharmacology; RNA, Messenger - genetics; Transcription (Genetics); Transcription, Genetic - drug effects; Transcription. Transcription factor. Splicing. Rna processing; Transcriptional regulatory elements; Transfection; Cyclic AMP; Regulation(control); Inhibitor; Transcription; Protein kinase
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.2821622

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.