1.The catalytic domain of xPAK1 is sufficient to induce myosin II dependent in vivo cell fragmentation independently of other apoptotic events.
Bisson N1, Islam N, Poitras L, Jean S, Bresnick A, Moss T. Dev Biol. 2003 Nov 15;263(2):264-81.
During apoptosis, cells are fragmented into sealed packages for safe disposal by phagocytosis, a process requiring major reorganisation of the cytoskeleton. The small p21 GTPase-activated kinases (PAKs) have been implicated in regulating cytoskeletal dynamics and a subset are activated by caspase 3/7 cleavage. However, the functional importance of this activation in apoptosis remains unknown. Using early Xenopus embryos, we have dissected xPAK1 activation from other causative events in apoptosis. An apoptotic-like cell fragmentation was observed 30 min after expression of the xPAK1 catalytic domain and occurred in the absence of other markers of apoptosis. In vitro, activated xPAK1 phosphorylated the regulatory light chain (xMLC) of myosin II at threonine 18 and serine 19, events known to activate the actin-dependent ATPase of cytoskeletal myosin. In vivo, activated xPAK1 induced hyperphosphorylation of xMLC. BDM, a myosin inhibitor, and ML-7, a MLCK inhibitor, both abrogated cell fragmentation induced by activated xPAK1, and ML-7 also inhibited xPAK1 activity.
2.Phospholipase C and myosin light chain kinase inhibition define a common step in actin regulation during cytokinesis.
Wong R1, Fabian L, Forer A, Brill JA. BMC Cell Biol. 2007 May 17;8:15.
BACKGROUND: Phosphatidylinositol 4,5-bisphosphate (PIP2) is required for successful completion of cytokinesis. In addition, both PIP2 and phosphoinositide-specific phospholipase C (PLC) have been localized to the cleavage furrow of dividing mammalian cells. PLC hydrolyzes PIP2 to yield diacylglycerol (DAG) and inositol trisphosphate (IP3), which in turn induces calcium (Ca2+) release from the ER. Several studies suggest PIP2 must be hydrolyzed continuously for continued cleavage furrow ingression. The majority of these studies employ the N-substituted maleimide U73122 as an inhibitor of PLC. However, the specificity of U73122 is unclear, as its active group closely resembles the non-specific alkylating agent N-ethylmaleimide (NEM). In addition, the pathway by which PIP2 regulates cytokinesis remains to be elucidated.
3.Regulatory effect of Rac1 on vascular reactivity after hemorrhagic shock in rats.
Li T1, Yang G, Xu J, Zhu Y, Liu L. J Cardiovasc Pharmacol. 2011 Jun;57(6):656-65. doi: 10.1097/FJC.0b013e318215e21d.
We used isolated superior mesenteric arteries (SMAs) from hemorrhagic-shock rats and hypoxia-treated vascular smooth muscle cells (VSMCs; mimicking the shock state) to observe the effects of platelet-derived growth factor (PDGF; Rac1 stimulator) and NSC23766 (Rac1 antagonist) on vascular reactivity and the relationship with the Rho kinase-myosin light-chain phosphatase (MLCP) and p21-activated kinase (PAK)-myosin light-chain kinase (MLCK) signal pathway. The results indicated that the contractile responses of the SMAs and VSMCs were significantly increased at early shock or after transient hypoxia. NSC23766 (Rac1 antagonist) further increased, whereas PDGF (Rac1 stimulator) decreased the contractile responses of SMAs and VSMCs. In the late period of shock or prolonged hypoxia, the contractile responses of SMAs and VSMCs were significantly decreased; NSC23766 increased (whereas PDGF further decreased) the contractile response of the SMAs and VSMCs.
4.Integrin-linked kinase is responsible for Ca2+-independent myosin diphosphorylation and contraction of vascular smooth muscle.
Wilson DP1, Sutherland C, Borman MA, Deng JT, Macdonald JA, Walsh MP. Biochem J. 2005 Dec 15;392(Pt 3):641-8.
Smooth muscle contraction is activated by phosphorylation at Ser-19 of LC20 (the 20 kDa light chains of myosin II) by Ca2+/calmodulin-dependent MLCK (myosin light-chain kinase). Diphosphorylation of LC20 at Ser-19 and Thr-18 is observed in smooth muscle tissues and cultured cells in response to various contractile stimuli, and in pathological circumstances associated with hypercontractility. MLCP (myosin light-chain phosphatase) inhibition can lead to LC20 diphosphorylation and Ca2+-independent contraction, which is not attributable to MLCK. Two kinases have emerged as candidates for Ca2+-independent LC20 diphosphorylation: ILK (integrin-linked kinase) and ZIPK (zipper-interacting protein kinase). Triton X-100-skinned rat caudal arterial smooth muscle was used to investigate the relative importance of ILK and ZIPK in Ca2+-independent, microcystin (phosphatase inhibitor)-induced LC20 diphosphorylation and contraction. Western blotting and in-gel kinase assays revealed that both kinases were retained in this preparation.
