We previously showed that ethanol regulates dopamine -hydroxylase (DBH) mRNA and protein levels in human neuroblastoma cells. DBH catalyzes norepinephrine synthesis and several studies have suggested a role for norepinephrine in ethanol-mediated behaviors. Here, we performed a detailed analysis of mechanism(s) underlying ethanol regulation of dbh expression in SH-SY5Y cells. Transient transfection analysis showed that ethanol (25-200 mM) caused concentration- and time-dependent increases in dbh gene transcription. Progressive deletions identified ethanol-responsive sequences in the -262 to -142 bp region of the dbh gene promoter. Mutagenesis of cAMP-response element (CRE) sequences in this region abolished ethanol-responsiveness, while maintaining responsiveness to phorbol esters. Co-expression of dominant-negative CREB greatly reduced ethanol induction of dbh. Inhibitors of protein kinase A, casein kinase II and MAP kinase reduced ethanol induction of dbh promoter activity. Pharmacogenomic studies with microarrays showed that PKA, MEK, or CKII inhibitors blocked induction of DBH and a large subset of ethanol-responsive genes. These genes had diverse functional groupings, including multiple members of the MAP kinase and phosphatidylinositol signaling cascades. Real-time PCR analysis validated select microarray results. Taken together, these results suggest that ethanol regulation of dbh requires a functional CRE element and its binding protein, CREB, and may require interaction of multiple kinase pathways. This mechanism may also mediate ethanol responsiveness of a complex subset of genes in neural cells. These studies improve our understanding of ethanol action at a cellular level and may have implications for behavioral responses to ethanol or mechanisms underlying ethanol-related neurological disease.