Inhibition of intracellular cAMP-dependent protein kinase using mutant genes of the regulatory type I subunit.
Expression vectors were constructed that code for mutated forms of the regulatory type 1 subunit (RI) of cyclic AMP-dependent protein kinase. These mutations alter a specific amino acid which is present in each of two homologous cAMP-binding domains of the RI protein. When these expression vectors were introduced into NIH 3T3 and Y1 adrenocortical tumor cells a mutant RI protein was produced that acted in a dominant fashion to cause a 20-400-fold inhibition of cAMP-dependent protein kinase activation. In addition, processes controlled by cAMP in adrenal cells were blocked; cells became resistant to the growth-inhibitory effects of cAMP and defective in steroid synthesis. Expression of mutant RI genes in cells provides a specific means to explore the role of cAMP and protein phosphorylation in the process of intracellular signalling.
Clegg
C H
CH
Department of Pharmacology, University of Washington, Seattle 98195.
Correll
L A
LA
Cadd
G G
GG
McKnight
G S
GS
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000502:physiology; D004789:Enzyme Activation; D005796:Genes; D005822:Genetic Vectors; D007700:Kinetics; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism
D046911:Macromolecular Substances; D047428:Protein Kinase Inhibitors; D000242:Cyclic AMP; D011494:Protein Kinases