Bryostatin 1 (Bryo), a macrocyclic lactone, stimulates some but not all of the biologic effects which are induced by phorbol esters (PEs). In vitro, it competes with PEs for binding to whole cells and activates the calcium/phospholipid-dependent protein kinase, PK-C. To examine whether Bryo, like PEs, is able to stimulate the nonadherent growth of cells, we used the mouse epidermal cell line JB6, which is stimulated by PEs to grow in soft agar. Like PEs, Bryo stimulates both the adherent and nonadherent growth of these cells, but Bryo (0.001-1 microM) is less active than equivalent concentrations of PEs. To attempt to explain the biologic differences between these two agents, we examined the modulation of PK-C by both PEs and Bryo. In a phosphotransferase assay using partially purified PK-C from JB6 cells, Bryo (1-0.001 microM) stimulated less phosphorylation of histone substrate than did PMA. Also, when whole cells were treated with equal concentrations of Bryo or PMA, Bryo stimulated a decreased loss of PK-C from the cytosol. Using purified isozymes of PK-C from rat brain, Bryo demonstrated identical competition to PMA for binding to forms alpha and gamma but decreased binding to form beta. Hydroxylapatite chromatography of JB6 cytosol demonstrated that these cells contain largely peak 2, or beta-PK-C. Although Bryo more weakly activates PK-C from JB6 cells, prolonged exposure of JB6 cells to either 1.0 or 0.01 microM Bryo caused a more rapid loss of immunologically detectable PK-C than did similar concentrations of PEs. We conclude that Bryo is capable of stimulating both the nonadherent and the adherent growth of JB6 cells in a similar fashion to phorbol esters. The differences in biologic effects of Bryo and PMA may be partially explained by Bryo's modulation of PK-C.