Interaction between Endothelial Differentiation-related Factor-1 and Calmodulin in Vitro and in Vivo *

Calmodulin (CaM) is the principal Ca²⁺ receptor protein inside the cell. When activated by Ca²⁺, CaM binds and activates target proteins, thus altering the metabolism and physiology of the cell. Under basal conditions, calcium-free CaM binds to other proteins termed CaM-binding proteins. Recently, we described endothelial differentiation-related factor (EDF)-1 as a protein involved in the repression of endothelial cell differentiation (Dragoni, I., Mariotti, M., Consalez, G. G., Soria, M., and Maier, J. A. M. (1998) J. Biol. Chem. 273, 31119–31124). Here we report that (i) EDF-1 binds CaMin vitro and in vivo; (ii) EDF-1 is phosphorylated in vitro and in vivo by protein kinase C; and (iii) EDF-1-CaM interaction is modulated by the concentrations of Ca²⁺ and by the phosphorylation of EDF-1 by protein kinase C both in vitro and in vivo. In addition, 12-O-tetradecanoylphorbol-13-acetate treatment of human umbilical vein endothelial cell stimulates the nuclear translocation of EDF-1. On the basis of the high homology of EDF-1 with multiprotein bridging factor-1, a transcriptional coactivator that binds TATA-binding protein (TBP), we also demonstrate that EDF-1 interacts with TBP in vitro and in human endothelial cells. We hypothesize that EDF-1 serves two main functions in endothelial cells as follows: (i) to bind CaM in the cytosol at physiologic concentrations of Ca²⁺ and (ii) to act in the nucleus as a transcriptional coactivator through its binding to TBP.