Phorbol esters and activators of protein kinase C have been reported to either facilitate or inhibit increases in intracellular cAMP caused by activators of adenylyl cyclase. The variable responses to activators of protein kinase C may reflect, in part, the existence of distinct adenylyl cyclases present in animal cells. There are a family of adenylyl cyclases with different regulatory properties, and clones for six distinct types of adenylyl cyclase have been reported. Two of these enzymes, the type I and type III adenylyl cyclases, are stimulated by calcium and calmodulin whereas the others are not. In this study, we examined the effect of phorbol esters of the activity of the type I and type III adenylyl cyclases in whole cells. TPA markedly enhanced the forskolin responsiveness of the type I and type III adenylyl cyclases expressed in kidney 293 cells. The effect of TPA on the activity of the calmodulin-sensitive adenylyl cyclases was not mediated through increases in intracellular free calcium. These data suggest that activation of protein kinase C can elevate intracellular cAMP in animal cells that contain the type I or type III adenylyl cyclase.