The expression of Wiskott-Aldrich syndrome protein (WASP) is dependent on WASP-interacting protein (WIP).

The Wiskott-Aldrich syndrome protein (WASP) is a key molecule for transduction of extracellular signals that induce a variety of critical biological events involving actin cytoskeleton rearrangement. Among the cellular partners of WASP, the Wiskott-Aldrich syndrome protein-interacting protein (WIP) has been speculated to play a critical role in the pathophysiology of Wiskott-Aldrich syndrome since WASP mutation hot spots map to the WIP-binding region. The notion that WIP promotes WASP function, however, conflicts with evidence that WIP inhibits WASP-mediated actin polymerization and IL-2 production and suggests a complex regulation of WASP function by WIP. Here we show that WASP gene transfer results in high WASP expression only when WIP is concomitantly expressed in K562 cells. Furthermore, WIP-knockdown experiments demonstrated that T cells with reduced WIP expression show a concordant reduction of WASP levels. Mapping studies using WIP mutants showed that the minimal WIP region able to rescue WASP expression in WIP-knockdown cells was the WASP-binding domain. However, expression of such a minimal domain of WIP failed to rescue WASP-dependent, nuclear factor of activated T-cells-mediated IL-2 transcriptional activity. These results demonstrate that expression of WIP is necessary for functional WASP expression in human cells and provide a new paradigm for understanding the function of these two molecules.

[1]  O. Danos,et al.  Lentiviral vectors targeting WASp expression to hematopoietic cells, efficiently transduce and correct cells from WAS patients , 2007, Gene Therapy.

[2]  U. Pannicke,et al.  Retroviral WASP gene transfer into human hematopoietic stem cells reconstitutes the actin cytoskeleton in myeloid progeny cells differentiated in vitro. , 2006, Experimental hematology.

[3]  L. Meng,et al.  WASP suppresses the growth defect of Saccharomyces cerevisiae las17Delta strain in the presence of WIP. , 2006, Biochemical and biophysical research communications.

[4]  L. Naldini,et al.  Efficacy of gene therapy for Wiskott-Aldrich syndrome using a WAS promoter/cDNA-containing lentiviral vector and nonlethal irradiation. , 2006, Human gene therapy.

[5]  Mariko Sawa,et al.  Caenorhabditis elegans WASP-interacting protein homologue WIP-1 is involved in morphogenesis through maintenance of WSP-1 protein levels. , 2006, Biochemical and biophysical research communications.

[6]  C. Frecha,et al.  Lentiviral vectors transcriptionally targeted to hematopoietic cells by WASP gene proximal promoter sequences , 2005, Gene Therapy.

[7]  P. Opolon,et al.  A lentiviral vector encoding the human Wiskott–Aldrich syndrome protein corrects immune and cytoskeletal defects in WASP knockout mice , 2005, Gene Therapy.

[8]  N. Caplen,et al.  Kinase-Independent Functions for Itk in TCR-Induced Regulation of Vav and the Actin Cytoskeleton1 , 2005, The Journal of Immunology.

[9]  Silvia Giliani,et al.  Mutations of the Wiskott-Aldrich Syndrome Protein (WASP): hotspots, effect on transcription, and translation and phenotype/genotype correlation. , 2004, Blood.

[10]  W. Vainchenker,et al.  Mechanisms of WASp-mediated hematologic and immunologic disease. , 2004, Blood.

[11]  T. Laín de Lera,et al.  Lentiviral vector-mediated gene transfer in T cells from Wiskott-Aldrich syndrome patients leads to functional correction. , 2004, Molecular therapy : the journal of the American Society of Gene Therapy.

[12]  F. Candotti,et al.  Multiple patients with revertant mosaicism in a single Wiskott-Aldrich syndrome family. , 2004, Blood.

[13]  J. Hartwig,et al.  WIP Regulates Signaling via the High Affinity Receptor for Immunoglobulin E in Mast Cells , 2004, The Journal of experimental medicine.

[14]  Kohsuke Imai,et al.  Clinical course of patients with WASP gene mutations. , 2004, Blood.

[15]  F. Candotti,et al.  Differential contribution of Wiskott-Aldrich syndrome protein to selective advantage in T- and B-cell lineages. , 2004, Blood.

[16]  H. Ochs.  The Wiskott-Aldrich syndrome , 1998, Springer Seminars in Immunopathology.

[17]  P. Doherty,et al.  Defects in T-cell-mediated immunity to influenza virus in murine Wiskott-Aldrich syndrome are corrected by oncoretroviral vector-mediated gene transfer into repopulating hematopoietic cells. , 2003, Blood.

[18]  L. Notarangelo,et al.  Wiskott-Aldrich Syndrome: a model for defective actin reorganization, cell trafficking and synapse formation. , 2003, Current opinion in immunology.

[19]  F. Candotti,et al.  Autoimmunity in Wiskott-Aldrich syndrome , 2003, Current opinion in rheumatology.

[20]  Michael J. Byrne,et al.  WIP participates in actin reorganization and ruffle formation induced by PDGF , 2003, Journal of Cell Science.

[21]  A. Nienhuis,et al.  Functional correction of T cells derived from patients with the Wiskott–Aldrich syndrome (WAS) by transduction with an oncoretroviral vector encoding the WAS protein , 2003, Gene Therapy.

[22]  F. Candotti,et al.  Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings. , 2003, The Journal of clinical investigation.

[23]  K. Siminovitch,et al.  The Wiskott–Aldrich syndrome protein: forging the link between actin and cell activation , 2003, Immunological reviews.

[24]  F. Alt,et al.  Gene therapy for Wiskott-Aldrich syndrome: rescue of T-cell signaling and amelioration of colitis upon transplantation of retrovirally transduced hematopoietic stem cells in mice. , 2003, Blood.

[25]  Alissa M. Weaver,et al.  Cortactin Interacts with WIP in Regulating Arp2/3 Activation and Membrane Protrusion , 2003, Current Biology.

[26]  Michael T. McManus,et al.  A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference , 2003, Nature Genetics.

[27]  M. Gandhi,et al.  X-linked thrombocytopenia caused by a mutation in the Wiskott-Aldrich syndrome (WAS) gene that disrupts interaction with the WAS protein (WASP)-interacting protein (WIP). , 2003, Experimental hematology.

[28]  Michael J. Byrne,et al.  Mechanism of recruitment of WASP to the immunological synapse and of its activation following TCR ligation. , 2002, Molecular cell.

[29]  W. Lim,et al.  Structure of the N-WASP EVH1 Domain-WIP Complex Insight into the Molecular Basis of Wiskott-Aldrich Syndrome , 2002, Cell.

[30]  Adrian J. Thrasher,et al.  Wasp in immune-system organization and function , 2002, Nature Reviews Immunology.

[31]  L. Dupré,et al.  Wiskott-Aldrich syndrome protein regulates lipid raft dynamics during immunological synapse formation. , 2002, Immunity.

[32]  G. Dunn,et al.  Restoration of podosomes and chemotaxis in Wiskott-Aldrich syndrome macrophages following induced expression of WASp. , 2002, The international journal of biochemistry & cell biology.

[33]  F. Candotti,et al.  Retrovirus-mediated WASP gene transfer corrects Wiskott-Aldrich syndrome T-cell dysfunction. , 2002, Human gene therapy.

[34]  J. Hartwig,et al.  WIP deficiency reveals a differential role for WIP and the actin cytoskeleton in T and B cell activation. , 2002, Immunity.

[35]  T. Takenawa,et al.  WASP and N-WASP in human platelets differ in sensitivity to protease calpain. , 2001, Blood.

[36]  D. Nelson,et al.  Somatic mosaicism in Wiskott–Aldrich syndrome suggests in vivo reversion by a DNA slippage mechanism , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[37]  A. Abo,et al.  A Role for Wiskott-Aldrich Syndrome Protein in T-cell Receptor-mediated Transcriptional Activation Independent of Actin Polymerization* , 2001, The Journal of Biological Chemistry.

[38]  J. Hartwig,et al.  WIP regulates N-WASP-mediated actin polymerization and filopodium formation , 2001, Nature Cell Biology.

[39]  D. Haas,et al.  Critical factors influencing stable transduction of human CD34(+) cells with HIV-1-derived lentiviral vectors. , 2000, Molecular therapy : the journal of the American Society of Gene Therapy.

[40]  P. Leibson,et al.  Cutting Edge: WIP, a Binding Partner for Wiskott-Aldrich Syndrome Protein, Cooperates with Vav in the Regulation of T Cell Activation1 , 2000, The Journal of Immunology.

[41]  Michael K. Rosen,et al.  Autoinhibition and activation mechanisms of the Wiskott–Aldrich syndrome protein , 2000, Nature.

[42]  S. Tsuboi,et al.  Expression of human Wiskott–Aldrich syndrome protein in patients’ cells leads to partial correction of a phenotypic abnormality of cell surface glycoproteins , 2000, Gene Therapy.

[43]  K. Siminovitch,et al.  Antigen Receptor–Induced Activation and Cytoskeletal Rearrangement Are Impaired in Wiskott-Aldrich Syndrome Protein–Deficient Lymphocytes , 1999, The Journal of experimental medicine.

[44]  F. Facchetti,et al.  Retrovirus-mediated WASP gene transfer corrects defective actin polymerization in B cell lines from Wiskott–Aldrich syndrome patients carrying ‘null’ mutations , 1999, Gene Therapy.

[45]  Matthew D. Welch,et al.  The Wiskott–Aldrich syndrome protein directs actin-based motility by stimulating actin nucleation with the Arp2/3 complex , 1999, Current Biology.

[46]  D. Nelson,et al.  Mutations that cause the Wiskott-Aldrich syndrome impair the interaction of Wiskott-Aldrich syndrome protein (WASP) with WASP interacting protein. , 1999, Journal of immunology.

[47]  Laura M. Machesky,et al.  Scar1 and the related Wiskott–Aldrich syndrome protein, WASP, regulate the actin cytoskeleton through the Arp2/3 complex , 1998, Current Biology.

[48]  R. Geha,et al.  The Wiskott-Aldrich Syndrome Protein-interacting Protein (WIP) Binds to the Adaptor Protein Nck* , 1998, The Journal of Biological Chemistry.

[49]  C. Kinnon,et al.  Absence of expression of the Wiskott-Aldrich syndrome protein in peripheral blood cells of Wiskott-Aldrich syndrome patients. , 1998, Clinical immunology and immunopathology.

[50]  Philip R. Cohen,et al.  Wiskott-Aldrich syndrome protein-deficient mice reveal a role for WASP in T but not B cell activation. , 1998, Immunity.

[51]  J. Hartwig,et al.  WIP, a protein associated with wiskott-aldrich syndrome protein, induces actin polymerization and redistribution in lymphoid cells. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[52]  O. Majdic,et al.  Expression of Wiskott-Aldrich syndrome protein (WASP) gene during hematopoietic differentiation. , 1997, Blood.

[53]  K. Miura,et al.  N‐WASP, a novel actin‐depolymerizing protein, regulates the cortical cytoskeletal rearrangement in a PIP2‐dependent manner downstream of tyrosine kinases. , 1996, The EMBO journal.

[54]  L. Notarangelo,et al.  Studies of the expression of the Wiskott-Aldrich syndrome protein. , 1996, The Journal of clinical investigation.

[55]  U. Francke,et al.  Wiskott–Aldrich Syndrome Protein, a Novel Effector for the GTPase CDC42Hs, Is Implicated in Actin Polymerization , 1996, Cell.

[56]  U. Francke,et al.  Isolation of a novel gene mutated in Wiskott-Aldrich syndrome , 1994, Cell.