The role of protein kinases in maintaining basal expression of voltage-gated K+ channel mRNA was examined in GH3 clonal pituitary cells. Nonspecific inhibition of protein kinases with H7 or staurosporine markedly decreases Kv1.5 K+ channel gene transcription and mRNA without producing a substantial change in Kv1.4 mRNA. Selective inhibitors for protein kinase C, Ca(2+)-calmodulin kinases, and tyrosine kinases do not affect Kv1.5 mRNA expression. In contrast, the Rp-diastereomer of adenosine 3',5'-cyclic monophosphorothioate, a specific inhibitor of protein kinase A, partially inhibits Kv1.5 mRNA expression (approximately 40%), and this effect was antagonized by 8-bromo-adenosine 3',5'-cyclic monophosphate. Thus, protein kinase A and at least one other kinase are required for basal Kv1.5 mRNA expression in pituitary cells.