The Wiskott-Aldrich Syndrome Protein-interacting Protein (WIP) Binds to the Adaptor Protein Nck*
- From the Division of ‡Immunology and ¶Howard Hughes Medical Institute, Children’s Hospital, Harvard Medical School, Boston, Massachusetts 02115
Abstract
Nck is a ubiquitous adaptor molecule composed of three Src homology 3 (SH3) domains followed by a single SH2 domain. Nck links, via its SH2 domain, tyrosine-phosphorylated receptors to effector proteins that contain SH3-binding proline-rich sequences. In this report, we demonstrate that recombinant Nck precipitates endogenous WIP, a novel proline-rich protein that interacts with the Wiskott-Aldrich syndrome protein (WASP), from BJAB cell lysates. Nck binds through its second SH3 domain to WIP, and Nck binds to WIP at a site (amino acids 321–415) that differs from the WASP-binding site (amino acids 416–488). WIP has been shown to associate with the actin polymerization regulatory protein profilin and to induce actin polymerization and cytoskeletal reorganization in lymphoid cells. We demonstrate the presence of profilin in Nck precipitates suggesting that Nck may couple extracellular signals to the cytoskeleton via its interaction with WIP and profilin.
Footnotes
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↵* This work was supported in part by National Institutes of Health Grants HL-59561-01 and AI-37130 (to N. R.) and by grants from the Baxter, Olsten, and Alpha Therapeutics Corp.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵§ Supported by a fellowship from Fundación Ramón Areces, Madrid, Spain.
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↵‖ To whom correspondence should be addressed: Enders 8, Division of Immunology, 300 Longwood Ave., Children’s Hospital, Boston, MA 02115. Tel.: 617-355-7602; Fax: 617-355-8205; E-mail:geha{at}a1.tch.harvard.edu.
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↵2 N. M. Quiles, I. M. Antón, N. Ramesh, and R. S. Geha, unpublished results.
- Abbreviations:
- SH3
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Src homology 3
- WASP
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Wiskott-Aldrich syndrome protein
- WIP
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WASP-interacting protein
- RTK
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receptor tyrosine kinase
- GST
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glutathioneS-transferase
- mAb
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monoclonal antibody
- PAGE
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polyacrylamide gel electrophoresis
- ECL
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enhanced chemiluminescent detection.
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- Received February 13, 1998.
- Revision received June 11, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
