Sphingosine 1-phosphate dynamically regulates myosin light chain phosphatase activity in human endothelial cells

Sphingosine 1-phosphate (SIP) is known to induce reorganization of the actin cytoskeleton through activation of the GTPase Rho. We have investigated the dynamic behavior of Rho/Rho kinase-regulated myosin light chain (MLC) phosphatase activity and MLC phosphorylation in Human Umbilical Vein Endothelial Cells (HUVEC) stimulated with SIP. Immediately (30-60 s) after SIP stimulation, MLC phosphatase activity dropped and MLC phosphorylation increased in a Rho/Rho kinase-dependent manner. Shortly thereafter (2 min), MLC phosphatase increased above baseline and MLC phosphorylation correspondingly decreased to near control values. At this time point formation of actin ruffles and Rac activity assays indicated activation of Rac. Finally, between 5 and 15 min, MLC phosphatase dropped to a plateau below baseline. In parallel, MLC phosphorylation became constantly elevated above control values. These findings indicate that SIP is able to induce dynamic cycles of MLC phosphatase deactivation and activation. This novel feature of SIP could contribute to its chemotactic and angiogenic activity. (C) 2002 Elsevier Science Inc. All rights reserved.