Stimulus-induced transcription of the Period gene is a critical step for phase-shift of vertebrate circadian systems. The promoter region of chicken Period2 contains a canonical calcium/cAMP-responsive element, but its functional relevance is not known. The present study shows that cAMP-responsive element-binding protein (CREB) and activating transcription factor-2 (ATF-2) bind to the promoter region of the Period2 gene in the chick pineal gland. In transient transfection assays, a reporter construct containing 0.7-kbp upstream region of chicken Period2 was transactivated by ATF-2, but it was poorly responsive to CREB. In the chick pineal gland, phosphorylation of CREB protein at the kinase-inducible domain was negatively regulated by light. On the other hand, phosphorylation of ATF-2 at the amino-terminal transactivation domain exhibited a circadian rhythm with a daytime peak, suggesting a role for ATF-2 in circadian rhythmicity in the chick pineal gland.