Dopamine beta-hydroxylase (DBH) catalyzes the conversion of dopamine to norepinephrine, and is expressed specifically in neurons and neuroendocrine cells that release norepinephrine and epinephrine. In the present study, we used DBH-expressing human neuroblastoma SK-N-BE(2)C and rat pheochromocytoma (PC12) cell lines to investigate the role of cAMP-dependent protein kinase (PKA) in transcriptional regulation of the DBH gene. Coexpression of the catalytic subunit of PKA (PKAc) robustly stimulated the transcriptional activity of the DBH gene in a dose-dependent manner. Conversely, coexpression of a specific inhibitor of PKA abrogated forskolin- and cAMP-mediated but not phorbol ester-mediated transcriptional induction of DBH. Deletion of the cAMP response element (CRE) dramatically reduced the stimulatory effect of PKA, indicating that the CRE mediates the induction of DBH by PKA. In DBH-nonexpressing HeLa and C6 glioma cell lines, coexpression of PKAc changed the transcriptional activity of the DBH promoter to a minimal degree, indicating that basal and PKA-mediated transcription of the DBH gene occur in a cell type-specific manner. Finally, both basal and cAMP-stimulated transcription of the DBH gene are diminished in three PKA-deficient PC12 cell lines, compared to wild-type cells. Based on these data, we conclude that PKA, via the CRE, plays an important role in basal and cAMP-inducible transcription, but is not required for phorbol ester-mediated induction, of the DBH gene in noradrenergic cells.(ABSTRACT TRUNCATED AT 250 WORDS)