ROLE OF VINCULIN IN EPITHELIAL CELL JUNCTIONS
Kris A Demali, Assistant Professor
University Of Iowa, Iowa City, Ia 52242
Grant 5K01CA111818-06 from National Cancer Institute
Abstract: The long-term goal of this research is to investigate the cytoskeletal regulation of cell-cell and cell-matrix adhesion-mediated events and the mechanism by which this regulation is subverted in neoplastic disease. The adhesion of cells to one another Is essential for the establishment and maintenance of normal tissue architecture. As such cell-cell contacts not only have an essential function in normal cell growth and motility but are also involved in cases of dysregulation that occur during carcinogenesis and metastasis. Cells have several mechanisms for linking to their neighbors. These so-called "cell junctions" consist of proteins that assemble into a variety of structures such as gap junctions, tight junctions, adherens junctions, desmosomes and hemidesmosomes, each with a different function. Despite their apparent stability, epithelial cell-cell junctions are highly dynamic regions of a cell that respond to cue from the extracellular environment. The morphological changes accompanying the response to these cues are mediated by the underlying actin cytoskeleton and their characteristics and integrity is controlled by kinases, phosphatases and small GTPases. A prominent phosphorylated protein that is reported to link cell-cell junctions to the actin cytoskeleton is vinculin. The role of vinculin at sites of adhesion to the matrix has been widely studied for many years, but the function of vinculin at sites of cell-cell contact remains largely unexplored. This proposal is directed towards answering this question. The hypothesis is that vinculin is an important regulator of epithelial cell junctions. This hypothesis will be tested and the role of vinculin dynamics and its phosphorylation in adherens and tight junction function will be examined
Keywords: Actins; Address; Adherens Junction; Adhering Junction; Adhesions; Adhesive Junction; Anchoring Junction; Architecture; Area; Binding; Binding (Molecular Function); Biochemical; Cancer Induction; Cell Adhesion; Cell Attachment; Cell Junctions; Cell Line; Cell Lines, Strains; Cell Locomotion; Cell Migration; Cell Movement; Cell-Cell Adhesion; Cell-Matrix Adhesions; Cell-Matrix Junction; CellLine; Cells; Cellular Adhesion; Cellular Expansion; Cellular Growth; Cellular Matrix; Cellular Migration; Characteristics; Chickens; Communicating Junction; Cues; Cytoskeletal System; Cytoskeleton; Desmosomes; Disease; Disorder; EC 2.7; Engineering / Architecture; Environment; Epithelial Cell Junction; Epithelial Cells; Event; Gallus domesticus; Gallus gallus; Gallus gallus domesticus; Gap Junctions; Goals; Hemidesmosomes; Human; Human, General; Intercellular Junctions; Invaded; Kinases; L-Serine; L-Threonine; Link; Low-resistance Junction; Macula Adherens; Maintenance; Maintenances; Man (Taxonomy); Man, Modern; Mediating; Metastasis; Metastasize; Metastatic Neoplasm; Metastatic Tumor; Modification; Molecular Interaction; Monomeric G-Proteins; Monomeric GTP-Binding Proteins; Motility; Motility, Cellular; Neoplasm Metastasis; Nexus; Nexus Junction; Normal Cell; Normal Tissue; Normal tissue morphology; Occluding Junctions; Phosphatases; Phosphohydrolases; Phosphomonoesterases; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Property; Property, LOINC Axis 2; Protein Phosphorylation; Proteins; RNA, Small Interfering; Regulation; Reporting; Research; Role; Secondary Neoplasm; Secondary Tumor; Serine; Shigella; Shigella Dysentery; Shigella Infections; Site; Small G-Proteins; Small GTPases; Small Interfering RNA; Spot Desmosome; Structure; Testing; Threonine; Tight Junctions; Transphosphorylases; Tumor Cell Invasion; Tumor Cell Migration; Tumor Invasion; Tyrosine Phosphorylation; VCL; Vinculin; Vinculin (Caenorhabditis elegans clone pRB9.1 protein moiety reduced); Zonula Occludens; cancer metastasis; carcinogenesis; cell growth; cell motility; cultured cell line; disease/disorder; domain mapping; extracellular; gene product; intracellular skeleton; mutant; neoplastic; pathogen; protein protein interaction; response; shigellosis; siRNA; social role
Project start date: 2005-08-19
Project end date: 2010-07-31
Budget start date: 1-AUG-2009
Budget end date: 31-JUL-2010
PFA/PA: PAR-03-104
5K01CA111818-06 (2009): $153900
Sponsored Links Excellgen http://Excellgen.com
Role Of Vinculin In Epithelial Cell Junctions
Kris A Demali
Biochemistryuniversity Of Iowa
iowa City, Ia 52242
Grant 5K01CA111818-05 from National Cancer Institute IRG: NCI
Abstract: The long-term goal of this research is to investigate the cytoskeletal regulation of cell-cell and cell-matrix adhesion-mediated events and the mechanism by which this regulation is subverted in neoplastic disease. The adhesion of cells to one another Is essential for the establishment and maintenance of normal tissue architecture. As such cell-cell contacts not only have an essential function in normal cell growth and motility but are also involved in cases of dysregulation that occur during carcinogenesis and metastasis. Cells have several mechanisms for linking to their neighbors. These so-called "cell junctions" consist of proteins that assemble into a variety of structures such as gap junctions, tight junctions, adherens junctions, desmosomes and hemidesmosomes, each with a different function. Despite their apparent stability, epithelial cell-cell junctions are highly dynamic regions of a cell that respond to cue from the extracellular environment. The morphological changes accompanying the response to these cues are mediated by the underlying actin cytoskeleton and their characteristics and integrity is controlled by kinases, phosphatases and small GTPases. A prominent phosphorylated protein that is reported to link cell-cell junctions to the actin cytoskeleton is vinculin. The role of vinculin at sites of adhesion to the matrix has been widely studied for many years, but the function of vinculin at sites of cell-cell contact remains largely unexplored. This proposal is directed towards answering this question. The hypothesis is that vinculin is an important regulator of epithelial cell junctions. This hypothesis will be tested and the role of vinculin dynamics and its phosphorylation in adherens and tight junction function will be examined
Project start date: 2005-08-19
Project end date: 2010-07-31
5K01CA111818-05 (2008): $153900
5K01CA111818-04 (2007): $153900
5K01CA111818-03 (2006): $132300
Grants awarded to Kris A Demali
Role Of Vinculin In Epithelial Cell Junctions
Kris A Demali
University Of North Carolina Chapel Hill Office Of Sponsored Research Chapel Hill, Nc 27599
Grant 1K01CA111818-01A1 from National Cancer Institute IRG: NCI
Abstract: The long-term goal of this research is to investigate the cytoskeletal regulation of cell-cell and cell-matrix adhesion-mediated events and the mechanism by which this regulation is subverted in neoplastic disease. The adhesion of cells to one another Is essential for the establishment and maintenance of normal tissue architecture. As such cell-cell contacts not only have an essential function in normal cell growth and motility but are also involved in cases of dysregulation that occur during carcinogenesis and metastasis. Cells have several mechanisms for linking to their neighbors. These so-called "cell junctions" consist of proteins that assemble into a variety of structures such as gap junctions, tight junctions, adherens junctions, desmosomes and hemidesmosomes, each with a different function. Despite their apparent stability, epithelial cell-cell junctions are highly dynamic regions of a cell that respond to cue from the extracellular environment. The morphological changes accompanying the response to these cues are mediated by the underlying actin cytoskeleton and their characteristics and integrity is controlled by kinases, phosphatases and small GTPases. A prominent phosphorylated protein that is reported to link cell-cell junctions to the actin cytoskeleton is vinculin. The role of vinculin at sites of adhesion to the matrix has been widely studied for many years, but the function of vinculin at sites of cell-cell contact remains largely unexplored. This proposal is directed towards answering this question. The hypothesis is that vinculin is an important regulator of epithelial cell junctions. This hypothesis will be tested and the role of vinculin dynamics and its phosphorylation in adherens and tight junction function will be examined.
Project start date: 2005-08-19
Project end date: 2005-12-31
1K01CA111818-01A1 (2005): $130140
REGULATION OF THE ARP2/3 COMPLEX BY INTEGRINS
Kris A Demali
Comprehensive Cancer Centeruniversity Of North Carolina Chapel Hill
office Of Sponsored Research
chapel Hill, Nc 27599
Grant 5F32GM020610-02 from National Institute Of General Medical Sciences IRG: CDF
Abstract: Adhesion to the extracellular matrix via integrins triggers cell spreading, and extension of filopodia and lamellipodia. A driving force for the extension of filopodia and lamellipodia is the polymerization of actin. Act- in polymerization can be initiated by either of two mechanisms by de novo polymerization, or the uncapping of capped actin filaments to expose the "barbed" end of the filaments, onto which additional actin subunits can polymerize. Current evidence indicates that the Arp2/3 complex plays a critical role in nucleating de novo polymerization, and that the Arp2/3 complex is involved in the actin polymerization that occurs during extension of lamellipodia and filopodia. Since integrin engagement promotes the formation of filopodia and lamellipodia, it seems likely therefore that there is a link between integrins and the Arp2/3 complex. The focus of this proposal is to determine if such a link exists and the nature of it. I would like to determine if Arp2/3 function contributes to the integrin- mediated cytoskeletal events, whether the Arp2/3 complex is recruited to the integrins, and if the Arp2/3 complex is tyrosine phosphorylated in response to integrin engagement. My hypothesis is that association of the Arp2/3 complex with integrins and tyrosine phosphorylation is an important determinant of the ability of cells to undergo events that are essential for cell movement and tumor cell invasion. Hence, these studies will provide novel information regarding how cell adhesion and actin polymerization are coordinated to give rise to cell motility and will yield information that will allow for a better understanding of the genesis and progression of tumor cell invasion and metastasis
Keywords: actin binding protein, integrin, polymerization antireceptor antibody, biological signal transduction, cell adhesion, cell migration, receptor binding 3T3 cell, antisense nucleic acid, confocal scanning microscopy, immunomagnetic separation, video recording system
5F32GM020610-02 (2001): $40196
1F32GM020610-01 (2000): $32416