Wiskott-Aldrich Syndrome (WAS) is an X-linked immunodeficiency disorder with dermatologic, hematologic and immunologic manifestations. The WAS gene ha been identified and, except for its association with Nck and Cdc42, very little is known about the biological function of its protein product WASP. Elucidation of the function of WASP is critical for understanding the molecular basis of the pathology of WAS. Using the yeast two hybrid system we have identified a novel gene, WIP (WASP- Interacting Protein), whose protein products interact with WASP. WIP displays homology to the yeast protein verprolin which is involved in maintaining the integrity of the cytoskeleton. Preliminary studies show that WIP associates with Nck. More importantly, overexpression of a truncated form of WIP containing the WASP-interacting domain causes disruption of actin stress fibers in CHO cells and a decrease in the microvilli in the human B cell line BJAB lymphocytes. We propose to: (1) Characterize WIP and perform a structural analysis of WIP-WASP interaction. We will a) determine the cellular localization of WIP; b) identify the domains and amino acid residues of WIP and WASP that are important for their interaction; c) investigate the association of WIP with WASP, Nck and Cdc42 in a multimeric complex and e) examine the interaction of WIP and WASP-related proteins. (2) Examine the structure and function of WIP with the cytoskeleton. We will a) examine the effect of overexpression of WIP and WIP mutants on the cytoskeleton; b) investigate the interaction of WIP with cytoskeletal proteins and c) determine if WIP complements verprolin deficient yeast. (3) Examine the role of WIP in signal transduction. We will a) analyze WIP phosphorylation and b) examine the interaction with rho GTPases, and the Nck associated enzymes PAK65 and PRK2. (4) Generate and analyze mice deficient in WIP. We will a) determine the organization of the murine WIP gene, b) generate and analyze WIP deficient mice. The results of the proposed studies should provide insights into the function of WIP and into the molecular pathology of the Wiskott-Aldrich syndrome.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL059561-04
Application #
6346234
Study Section
Project Start
2000-09-01
Project End
2001-08-31
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
4
Fiscal Year
2000
Total Cost
$426,552
Indirect Cost
Name
Immune Disease Institute, Inc.
Department
Type
DUNS #
115524410
City
Boston
State
MA
Country
United States
Zip Code
02115
Garber, John J; Mallick, Emily M; Scanlon, Karen M et al. (2018) Attaching-and-Effacing Pathogens Exploit Junction Regulatory Activities of N-WASP and SNX9 to Disrupt the Intestinal Barrier. Cell Mol Gastroenterol Hepatol 5:273-288
Vardi, Iddo; Barel, Ortal; Sperber, Michal et al. (2018) Genetic and Structural Analysis of a SKIV2L Mutation Causing Tricho-hepato-enteric Syndrome. Dig Dis Sci 63:1192-1199
Li, Jian; Shouval, Dror S; Doty, Andria L et al. (2017) Increased Mucosal IL-22 Production of an IL-10RA Mutation Patient Following Anakin Treatment Suggests Further Mechanism for Mucosal Healing. J Clin Immunol 37:104-107
Lexmond, Willem S; Goettel, Jeremy A; Lyons, Jonathan J et al. (2016) FOXP3+ Tregs require WASP to restrain Th2-mediated food allergy. J Clin Invest 126:4030-4044
Baptista, Marisa A P; Keszei, Marton; Oliveira, Mariana et al. (2016) Deletion of Wiskott-Aldrich syndrome protein triggers Rac2 activity and increased cross-presentation by dendritic cells. Nat Commun 7:12175
Volpi, Stefano; Santori, Elettra; Abernethy, Katrina et al. (2016) N-WASP is required for B-cell-mediated autoimmunity in Wiskott-Aldrich syndrome. Blood 127:216-20
Moran, Christopher J; Klein, Christoph; Muise, Aleixo M et al. (2015) Very early-onset inflammatory bowel disease: gaining insight through focused discovery. Inflamm Bowel Dis 21:1166-75
Crestani, Elena; Volpi, Stefano; Candotti, Fabio et al. (2015) Broad spectrum of autoantibodies in patients with Wiskott-Aldrich syndrome and X-linked thrombocytopenia. J Allergy Clin Immunol 136:1401-4.e1-3
Kolhatkar, Nikita S; Brahmandam, Archana; Thouvenel, Christopher D et al. (2015) Altered BCR and TLR signals promote enhanced positive selection of autoreactive transitional B cells in Wiskott-Aldrich syndrome. J Exp Med 212:1663-77
Gerasimcik, Natalija; Dahlberg, Carin I M; Baptista, Marisa A P et al. (2015) The Rho GTPase Cdc42 Is Essential for the Activation and Function of Mature B Cells. J Immunol 194:4750-8

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