The ₁-subunit of the cardiac/vascular Ca²⁺ channel, which is the dihydropyridine (DHP)-binding site (the DHP receptor), provides the pore structure for Ca²⁺ entry. It contains the binding sites for multiple classes of drugs collectively known as Ca²⁺ antagonists. As an initial step toward understanding the mechanisms controlling transcription of the rat cardiac _1C-subunit gene, we have cloned a 2.3-kb fragment containing the 5'-flanking sequences and identified the _1C-subunit gene transcription start site. The rat _1C-subunit gene promoter belongs to the TATA-less class of such basal elements. Using deletion analysis of _1C-subunit promoter-luciferase reporter gene constructs, we have characterized the transcriptional modulating activity of the 5'-flanking region and conducted transient transfections in cultured neonatal rat cardiac ventricular myocytes and vascular smooth muscle cells. Sequence scanning identified several potential regulatory elements, including five consensus sequences for the cardiac-specific transcription factor Nkx2.5, an AP-1 site, a cAMP response element, and a hormone response element. Transient transfection experiments with the promoter-luciferase reporter fusion gene demonstrate that the 2-kb 5'-flanking region confers tissue specificity and hormone responsiveness to expression of the Ca²⁺ channel _1C-subunit gene. Electrophoretic mobility shift assays identified a region of the _1C-subunit gene promoter that can bind transcription factors and appears to be important for gene expression.