Protrusive and contractile cell-matrix adhesions have important roles in cell adhesion, movement and signaling. Until recently, there has been a paucity of systems with which to research cell protrusions at molecular level. This laboratory previously discovered that thrombospondin-1, a cell-adhesion molecule, specifically induces protrusive lamellipodia, spikes and filopodia that contain the actin cross-linking protein, fascin, in core F-actin bundles. These fascin-containing structures are necessary for cell adhesion and migration on TSP-1 substrata, contribute to adhesion and migration on fibronectin substrata and are also formed in response to certain peptide growth factors. We have identified a novel molecular pathway by which thrombospondin-1 acts through clustering of the cell-surface proteoglycan syndecan-1 to stimulate the assembly of fascin-containing protrusions. The central hypothesis of the proposed research is that molecular components necessary for assembly of fascin-containing cell protrusions participate in protein complexes that contribute to the regulation of cell-matrix adhesion and cell movement. The proposed research focuses on the elucidatation of the molecular processes downstream of syndecan- 1 that mediate the organization of fascin cytoskeletal structures, the initial integration of these molecular processes into cell signaling networks, and the identification of mechanisms by which fascin-containing protrusions contribute to cell movements in three-dimensional matrix. These goals will be addressed experimentally by, 1), elucidation of the molecular requirements for regions within the syndecan-1 extracellular and cytoplasmic domains in transducing the organization of fascin protrusions; 2), the discovery of key binding partners for the V region of the syndecan-1 cytoplasmic domain and how these are integrated into regulation of fascin organization through the activation of Cdc42 and Rac small GTPases ; 3), the identification of molecular processes by which fascin structures contribute to migration in 3-dimensional matrix. Success in these complementary aims will advance new knowledge and understanding of the roles of fascin-based cytoskeletal structures in the important fundamental cellular activities of matrix-adhesion and cell migration. By opening up a new understanding of the molecular processes downstream of a specific extracellular inducer of fascin-containing protrusions, the proposed research will develop knowledge that is an essential basis for further investigation of fascin protrusions and associated regulatory molecules as therapeutic targets.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM068073-04
Application #
7072797
Study Section
Special Emphasis Panel (ZRG1-CDF-4 (02))
Program Officer
Deatherage, James F
Project Start
2003-06-01
Project End
2008-05-31
Budget Start
2006-06-01
Budget End
2007-05-31
Support Year
4
Fiscal Year
2006
Total Cost
$291,339
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
Kim, Dae Joong; Christofidou, Elena D; Keene, Douglas R et al. (2015) Intermolecular interactions of thrombospondins drive their accumulation in extracellular matrix. Mol Biol Cell 26:2640-54
Bentley, Amber A; Adams, Josephine C (2010) The evolution of thrombospondins and their ligand-binding activities. Mol Biol Evol 27:2187-97
Adams, Josephine C; Bentley, Amber A; Kvansakul, Marc et al. (2008) Extracellular matrix retention of thrombospondin 1 is controlled by its conserved C-terminal region. J Cell Sci 121:784-95
Valiyaveettil, Manojkumar; Bentley, Amber A; Gursahaney, Priya et al. (2008) Novel role of the muskelin-RanBP9 complex as a nucleocytoplasmic mediator of cell morphology regulation. J Cell Biol 182:727-39
Parsons, Maddy; Adams, Josephine C (2008) Rac regulates the interaction of fascin with protein kinase C in cell migration. J Cell Sci 121:2805-13
Chakravarti, Ritu; Adams, Josephine C (2006) Comparative genomics of the syndecans defines an ancestral genomic context associated with matrilins in vertebrates. BMC Genomics 7:83
Chakravarti, Ritu; Sapountzi, Vasileia; Adams, Josephine C (2005) Functional role of syndecan-1 cytoplasmic V region in lamellipodial spreading, actin bundling, and cell migration. Mol Biol Cell 16:3678-91
Adams, Josephine C (2004) Fascin protrusions in cell interactions. Trends Cardiovasc Med 14:221-6
Adams, Josephine C (2004) Roles of fascin in cell adhesion and motility. Curr Opin Cell Biol 16:590-6
De Arcangelis, Adele; Georges-Labouesse, Elisabeth; Adams, Josephine C (2004) Expression of fascin-1, the gene encoding the actin-bundling protein fascin-1, during mouse embryogenesis. Gene Expr Patterns 4:637-43