Secretin evokes catecholamine secretion from PC12 pheochromocytoma cells. We tested whether secretin activates transcription of the major vesicular core protein chromogranin A (CgA). Secretin stimulated both endogenous CgA gene transcription (approximately 4-6-fold) as well as transfected CgA promoter activity (approximately 8-10-fold; EC50, approximately 7 nm) in PC12 cells. Studies on CgA promoter 5'-deletion mutant/luciferase reporter constructs, point mutations of the CgA cAMP response element (CRE), and their transfer to a heterologous promoter implicated CRE in cis as both necessary and sufficient for secretin-stimulated CgA gene transcription. Secretin-induced CgA gene transcription was inhibited/abolished by cytosolic Ca2+ chelation, chemical blockade of phospholipase C, protein kinase A (PKA), or mitogen-activated protein (MAP) kinase extracellular signal regulated kinase (ERK) 1/2 and the expression of dominant negative mutants of ERK1/2, CRE binding protein (CREB) kinase RSK2, or CREB. Secretin also augmented (approximately 4-fold) phosphorylation of ERK1/2. Trans-activation (approximately 21-fold) of GAL4-CREB fusion protein by secretin indicates involvement of CREB in secretin signaling to gene transcription. Electrophoretic mobility shift assays also identified CREB as the mediator of secretin-induced CgA gene transcription, and pCREB supershifts indicated Ser-133 as the active CREB moiety in vitro. This conclusion was reinforced in vivo by results of chromatin pCREB immunoprecipitation assays. We conclude that secretin signals to CgA gene transcription through the CRE domain in cis and through cAMP, Ca2+, PKA, MAP kinase, and the transcription factor CREB in trans. Thus, multiple signal transduction pathways seem to subserve the function of stimulus-transcription coupling after this peptidergic stimulus to chromaffin cells.