|Budget Amount *help
¥2,800,000 (Direct Cost : ¥2,800,000)
Fiscal Year 1999 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 1998 : ¥2,000,000 (Direct Cost : ¥2,000,000)
We found Rho-associated coiled coil forming protein kinase (p160ROCK) is serine/threonine kinase. p160ROCK is a member of effector protein of small GTPase Rho and specisically binds to activated form of Rho thereby becomes activated upon this binding. During transfction experiments in fibroblast and eptherial cells, this kinase is involved in formation of actin stress fibers and focal adhesions mediated by Rho. In addition to this, in nuroblastama N1E-115 cell this kinase induces cell rounding by regulating contractile force at cell cortex.
Thus, it is thought that p160ROCK is localized to several specific sites in the cell and phosphorylates several substrates.
Recently, we developed a specific inhibitor of ROCK, we named it Y-27632. This compound competes with ATP at the kinase domain. Using this compound, we reported that Rho/ROCK pathway regulates blood pressure in hypertension model rats, indicating that this pathway is involved in the pathophysiology of hypertension. However, the molecular mechanism of Rho/ROCK pathway is not clearly known.
In this study, we try to understand the molecular mechanism (s) of p160ROCK-mediated signaling pathway (s) through the isolation of phosphorylated substrates for p160ROCK.
As a result, our major findings are as follows,
1. p160ROCK regulates Confilin/ADF phosphorylaytion through phosphorylation and activation of LIM-kinase.
2. p160ROCK cooperates with mDia, another effector molecule of Rho, on Rho-mediated actin stress fiber formation and focal adhesion formation.