The roles of cAMP and CAMP-dependent protein kinase in the regulation of adrenocortical functions: analysis using DNA-mediated gene transfer

Wong, M., O'Brien, D. & Schimmer, B. P. (1986) The roles of cAMP and CAMP-dependent protein kinase in the regulation of adrenocortical functions: analysis using DNA-mediated gene transfer. Biochem. Cell Biol. 64, 1066-1071 DNA-mediated gene transfer was used to evaluate the cause and effect relationship between mutations in CAMP-dependent protein kinase activity and cellular resistance of adrenocortical tumor cells to ACTH and CAMP. Protein kinase defective, Kin 8 adrenocortical tumor cells were transformed with genomic DNA from an ACTH- and CAMP-responsive adrenocortical cell line and screened for the recovery of morphological responses to the cAMP analog 8-bromo-CAMP (8BrcAMP). 8BrcAMPresponsive transformants were recovered with a frequency of approximately 0.5 per lo3 transformation-competent cells. These transformants recovered the ability to round up in the presence of ACTH and were able to respond to both ACTH and 8BrcAMP with increased steroidogenesis. They also recovered CAMP-dependent protein kinase activity. The transformants, however, were unstable and concomitantly lost CAMP-dependent protein kinase activity and steroidogenic and morphological responses to ACTH and 8BrcAMP. These observations suggest that a single gene, probably the gene encoding the regulatory subunit of CAMP-dependent protein kinase, is responsible for the resistance of the Kin 8 mutant to ACTH and CAMP. Wong, M., O'Brien, D. & Schimmer, B. P. (1986) The roles of cAMP and CAMP-dependent protein kinase in the regulation of