This proposal focuses on deciphering myosin II isoform roles and regulation during cytokinesis and during polarized cell migration and chemotactic responses. Although myosin II is a critical and ubiquitous force producing enzyme, its exact mechanical roles during cellular contractile events is poorly understood, and there is virtually no understanding of isoform specific roles. Multiple conflicting models exist regarding how myosin II filament assembly and localization is regulated in vivo, and none of the prevailing models have been tested in vivo. We will use in vivo imaging of GFP-myosin II reporter proteins, fluorescent recovery after photobleaching (FRAP) technology, as well as siRNA and other approaches to generate significant new understanding of fundamental mechanisms of myosin II regulation and new understanding of the isoform-specific roles of myosin MA and MB in processes such as cytokinesis and polarized cell migration. Our studies will also identify the key upstream protein kinases and regulatory proteins responsible for activation of myosin II assembly and contractility during cytokinesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM077224-05
Application #
7792304
Study Section
Special Emphasis Panel (ZRG1-NDT-K (01))
Program Officer
Deatherage, James F
Project Start
2007-04-01
Project End
2012-03-31
Budget Start
2010-04-01
Budget End
2012-03-31
Support Year
5
Fiscal Year
2010
Total Cost
$272,003
Indirect Cost
Name
Cleveland Clinic Lerner
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
135781701
City
Cleveland
State
OH
Country
United States
Zip Code
44195
Beach, Jordan R; Hussey, George S; Miller, Tyler E et al. (2011) Myosin II isoform switching mediates invasiveness after TGF-?-induced epithelial-mesenchymal transition. Proc Natl Acad Sci U S A 108:17991-6
Beach, Jordan R; Licate, Lucila S; Crish, James F et al. (2011) Analysis of the role of Ser1/Ser2/Thr9 phosphorylation on myosin II assembly and function in live cells. BMC Cell Biol 12:52
Betapudi, Venkaiah; Gokulrangan, Giridharan; Chance, Mark R et al. (2011) A proteomic study of myosin II motor proteins during tumor cell migration. J Mol Biol 407:673-86
Betapudi, Venkaiah; Rai, Vandana; Beach, Jordan R et al. (2010) Novel regulation and dynamics of myosin II activation during epidermal wound responses. Exp Cell Res 316:980-91
Betapudi, Venkaiah (2010) Myosin II motor proteins with different functions determine the fate of lamellipodia extension during cell spreading. PLoS One 5:e8560
Breckenridge, Mark T; Egelhoff, Thomas T; Baskaran, Harihara (2010) A microfluidic imaging chamber for the direct observation of chemotactic transmigration. Biomed Microdevices 12:543-53
Breckenridge, Mark T; Dulyaninova, Natalya G; Egelhoff, Thomas T (2009) Multiple regulatory steps control mammalian nonmuscle myosin II assembly in live cells. Mol Biol Cell 20:338-47
Beach, Jordan R; Egelhoff, Thomas T (2009) Myosin II recruitment during cytokinesis independent of centralspindlin-mediated phosphorylation. J Biol Chem 284:27377-83