Robert Wysolmerski, Ph.D.
	Professor of Neurobiology and Anatomy Graduate Training: St. Louis University Fellowship: St. Louis University
	Office: 211 BBRF Lab: 270 BBRF PO Box 9105 Morgantown, WV 26506	Email: rwysomerski@hsc.wvu.edu Phone: 304-293-2213 Fax: 304-293-5513

Research Interests:

The overall aim of the laboratory is to define the events that initiate and regulate endothelial cell contraction in an effort to elucidate the basis of increased vascular permeability. The primary hypothesis underlying the proposed work dictates that exposure to inflammatory mediators activates the endothelial cell actomyosin based contractile system. Calcium (Ca 2+) dependent and/or Ca 2+-independent stimulated phosphorylation of the myosin II regulatory light chain results in myosin II filament formation, an obligatory step leading to endothelial cell motility and contraction. In nonmuscle cells, myosin II is a major cytoskeletal protein which has the ability to convert the chemical energy of ATP into the mechanical work of cell contraction through its interaction with actin. In nonmuscle cells, MLCK-catalyzed phosphorylation results in mono- and diphosphorylation of regulatory light chains at site Ser19 or sites Ser19 and Thr18, respectively. In endothelial cells, we have shown that a small GTPase dependent enzyme, p-21 activated kinase (PAK2), catalyzes Ca 2+ independent activation of nonmuscle myosin II by phosphorylation of the regulatory light chain that is restricted to site Ser19. This PAK2-mediated monophosphorylation results in a less forceful contractile response than when the regulatory light chain is diphosphorylated by MLCK. More recent studies have shown that PAK2 also phosphorylates unactivated MLCK which results in inhibition of MLCK activation by Ca 2+. Collectively, these findings have clearly established a role for PAK2 in mediating and regulating myosin II activity, and thus contractile activity, in endothelial cells.

The goal of our studies is to biochemically and morphologically characterize myosin II activation by Ca 2+ -dependent and Ca 2+ -independent signaling pathways in endothelial cells The working hypothesis is that a specific pools of myosin II, determined by the heavy chain isoform specificity (myosin IIA vs IIB vs IIC), are activated by enzyme specific modifications. Functionally, regulated differential phosphorylation of myosin II would allow the endothelial cell to react to a variety of physiological signals with graded contractile responses.

Selected Publications:

• Blue, E. K., Goeckeler, Z. M., Jin, Y., Hou, L., Dixon,S. A., Herring, B. P., Wysolmerski, R. B. and Gallagher, P. J. 220 and 130 kDa MLCKs have distinct tissue distributions and intracellular localization patterns. Am. J. Physiol. 282: C451-C460, 2002.

• Thukkani, A. K., Hsu, F., Crowly, J. R., Wysolmerski, R. B., Albert, C. A. and Ford, D. A. Reactive chlorinating species produced during neutrophil activation target tissue plasmalogens. J. Biol. Chem. 277: 3242-3849, 2002.

• Krts, I., Wysolmerski, R. B., Holliday, S. and Lee, B.S. Distribution of myosin II isoforms in resting and activated osteoclasts. Calcif. Tissue Int. 71: 530-538, 2002.

• Wakatsuki, T., Wysolmerski, R. B. and Elson, E. L. Mechanics of cell spreading: role of myosin II. J Cell Science 116: 1617-1625, 2003.

• Martinson, B.D., Albert, C. J., Corbett, J. A., Wysolmerski, R.B. and Ford, D.A. Calcium-independent phospholipase A2 mediates CREB phosphorylation in double-stranded RNA-stimulated endothelial cells. J. Lipid Res 44: 1686-1691, 2003.

• Emmert, D.A., Fee, J. A., Goeckeler, Z. M., Wakatsuki, T., Herring, P., Gallagher, P., Elson, E. L. and Wysolmerski, R. B. Rho-kinase-mediated calcium-independent contraction in rat embryo fibroblasts. Am J Physiol Cell Physiol. 286: C8-C21, 2004.

• Goeckeler, Z.M., and Wysolmerski, R.B. Myosin phosphatase and cofilin mediate cAMP/cAMPdependent protein kinase-induced decline in endothelial cell isometric tension and myosin II regulatory light chain phosphorylation. J. Biol. Chem. 280: 33083-33095, 2005.

• Xu, M., Waters, C., Hu, C., Wysolmerski, R.B., Vincent, P.A., Minnear, F.L. Sphingosine 1-Phosphate increases endothelial electrical resistance independently of VE-cadherin but requires cell spreading and rho-kinase. Am J Physiol. Cell Physiol. 239: C1309-1318, 2007.

• Goeckeler, Z.M., Bridgman, P.C. and Wysolmerski, R.B. Nonmuscle Myosin II is Responsible for Maintaining Endothelial Cell Basal Tone and Stress Fiber Integrity. Am J Physiol. Cell Physiol: 295:C994-C1006, 2008.

• Brown, J., Wysolmerski, R.B. and Bridgman, P.C. A biphasic response of dorsal root ganglion to semaphoring 3A requires multiple myosin II isoforms. Mol. Biol Cell, 20:1167-1179, 2009.
  

Lab Personnel: Kimberly Miede -Ph.D. Student Jing Zhu - Ph.D. Student
	Kimberly Miede and Jing Zhu