title: Src kinase Hck association with the WASp and mDia1 cytoskeletal regulators promotes chemoattractant-induced Hck membrane targeting and activation in neutrophils.
abstract: The haemopoietic cell kinase (Hck) plays an important but poorly understood role in coupling chemoattractant stimuli to the actin cytoskeletal rearrangement required for neutrophil polarization and chemotaxis. Here, we show that Hck coimmunoprecipitates with the cytoskeletal regulatory Wiskott-Aldrich syndrome protein (WASp) and mammalian diaphanous-related formin 1 (mDia1) in chemoattractant-stimulated neutrophils, and that the 3 proteins inducibly colocalize with one another at the leading edge of chemotaxing cells. Hck interaction with WASp was found to be mediated by the Hck SH3 domain binding to the WASp proline-rich region, while Hck interaction with mDia1 was indirect but was required for binding to WASp. In contrast to wild-type cells, both WASp- and mDia1-deficient neutrophils showed severe impairment of chemokine-induced Hck membrane translocation and induction of Hck binding to WASp, and Hck activation and WASp tyrosine phosphorylation were impaired in mDia1-/- cells. Thus, chemotactic stimulation appears to induce an mDia1/Hck/WASp complex required for Hck membrane targeting and for induction of the Hck-mediated WASp tyrosine phosphorylation thought to be required for WASp-driven actin polymerization. These findings reveal that Hck functions in neutrophils to be realized, at least in part, via its interaction with mDia1 and WASp, and identifies the mDia1/Hck/WASp axis as a cytoskeletal signaling interface linking tyrosine phosphorylation to chemotactic and, possibly, other actin-based neutrophil responses.
authors.affiliation: Department of Medicine, Immunology, and Molecular and Medical Genetics, University of Toronto, Toronto, ONM5G1X5, Canada.
authors.forename: Yongquan
authors.initials: Y
authors.lastname: Shi
authors.forename: Baoxia
authors.initials: B
authors.lastname: Dong
authors.forename: Helen
authors.initials: H
authors.lastname: Miliotis
authors.forename: Junye
authors.initials: J
authors.lastname: Liu
authors.forename: Arthur S
authors.initials: AS
authors.lastname: Alberts
authors.forename: Jinyi
authors.initials: J
authors.lastname: Zhang
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Siminovitch
mesh_terms: D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology*; D018925:Chemokines / Q000494:pharmacology; D002630:Chemotactic Factors / Q000494:pharmacology*; D002633:Chemotaxis / Q000187:drug effects; D002522:Chlorocebus aethiops; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D000080865:Formins; D018922:HL-60 Cells; D006801:Humans; D051379:Mice; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D019000:Phosphotyrosine / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D051572:Proto-Oncogene Proteins c-hck / Q000378:metabolism*; D011554:Pseudopodia / Q000187:drug effects / Q000201:enzymology; D051316:Wiskott-Aldrich Syndrome Protein / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D018925:Chemokines; D002630:Chemotactic Factors; C491464:Diap1 protein, mouse; D000080865:Formins; D051316:Wiskott-Aldrich Syndrome Protein; D019000:Phosphotyrosine; C496632:Hck protein, mouse; D051572:Proto-Oncogene Proteins c-hck