Tyrosine phosphorylation of Wiskott-Aldrich Syndrome Protein (WASP) by Hck regulates macrophage function

Haein Park, Athanassios Dovas, Samer Hanna, Claire Lastrucci, Celine Cougoule, Romain Guiet, Isabelle Maridonneau-Parini, Dianne Cox

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13 Scopus citations

Abstract

We have shown previously that tyrosine phosphorylation of Wiskott-Aldrich syndrome protein (WASP) is important for diverse macrophage functions including phagocytosis, chemotaxis, podosome dynamics, and matrix degradation. However, the specific tyrosine kinase mediating WASP phosphorylation is still unclear. Here, we provide evidence that Hck, which is predominantly expressed in leukocytes, can tyrosine phosphorylate WASP and regulates WASP-mediated macrophage functions. We demonstrate that tyrosine phosphorylation of WASP in response to stimulation with CX3CL1or via Fcγ receptor ligation were severely reduced in Hck-/- bone marrow-derived macrophages (BMMs) or in RAW/LR5 macrophages in which Hck expression was silenced using RNA-mediated interference (Hck shRNA). Consistent with reduced WASP tyrosine phosphorylation, phagocytosis, chemotaxis, and matrix degradation are reduced in Hck -/- BMMs or Hck shRNA cells. In particular, WASP phosphorylation was primarily mediated by the p61 isoform of Hck. Our studies also show that Hck and WASP are required for passage through a dense three-dimensional matrix and transendothelial migration, suggesting that tyrosine phosphorylation of WASP by Hck may play a role in tissue infiltration of macrophages. Consistent with a role for this pathway in invasion, WASP-/- BMMs do not invade into tumor spheroids with the same efficiency as WT BMMs and cells expressing phospho-deficient WASP have reduced ability to promote carcinoma cell invasion. Altogether, our results indicate that tyrosine phosphorylation of WASP by Hck is required for proper macrophage functions.

Original languageEnglish (US)
Pages (from-to)7897-7906
Number of pages10
JournalJournal of Biological Chemistry
Volume289
Issue number11
DOIs
Publication statusPublished - Mar 14 2014

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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