Dopamine beta-hydroxylase (DBH), the enzyme catalyzing the conversion of dopamine to norepinephrine, is specifically expressed in adrenergic and noradrenergic neurons in the central nervous system. DNase I hypersensitive sites were found in the 5'-flanking region of the DBH gene in noradrenergic human neuroblastoma SK-N-BE(2)C cells, but not in DBH-nonexpressing HeLa cells. We report here that the 4.3-kilobase upstream sequence of the human DBH gene confers cell type-specific expression as assessed by transient expression assay. Furthermore, deletional and mutational analyses revealed two genetic regulatory elements required for the regulation of cell type specificity. First, deletion of the cAMP-response element (CRE) abolished > 95% of the transcriptional activity by the DBH upstream promoter, thus implicating the CRE as an essential positive genetic element. Second, deletion of a region between -490 and -263 base pairs resulted in 10-fold increase of reporter gene activity only in HeLa cells, indicating that this region contains a cell-specific silencer. A 13-base pair fragment residing within that region shows 77% sequence identity with the neuron-specific silencer motif recently identified in two neuronal genes, i.e. SCG10 and type II sodium channel genes. We propose that the interplay between the CRE and this neuron-specific silencer region plays an important role in the tissue-specific expression of the DBH gene in noradrenergic cells.