title: WASH has a critical role in NK cell cytotoxicity through Lck-mediated phosphorylation.
abstract: Natural killer (NK) cells are important effector cells of the innate immune system to kill certain virus-infected and transformed cells. Wiskott-Aldrich Syndrome protein (WASP) and SCAR homolog (WASH) has been identified as a member of WASP family proteins implicated in regulating the cytoskeletal reorganization, yet little is known about its function in lymphocytes. Here we demonstrate that WASH is crucial for NK cell cytotoxicity. WASH was found to colocalize with lytic granules upon NK cell activation. Knockdown of WASH expression substantially inhibited polarization and release of lytic granules to the immune synapse, resulting in the impairment of NK cell cytotoxicity. More importantly, our data also define a previously unappreciated mechanism for WASH function, in which Src family kinase Lck can interact with WASH and induce WASH phosphorylation. Mutation of tyrosine residue Y141, identified here as the major site of WASH phosphorylation, partially blocked WASH tyrosine phosphorylation and NK cell cytotoxicity. Taken together, these observations suggest that WASH has a pivotal role for regulation of NK cell cytotoxicity through Lck-mediated Y141 tyrosine phosphorylation.
authors.affiliation: Key Laboratory of Infection and Immunity of CAS, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.|Biotherapy Center, The First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: L
authors.initials: L
authors.lastname: Huang
authors.affiliation: Key Laboratory of Infection and Immunity of CAS, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: P
authors.initials: P
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Infection and Immunity of CAS, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: P
authors.initials: P
authors.lastname: Xia
authors.affiliation: Key Laboratory of Infection and Immunity of CAS, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Fan
mesh_terms: D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D002460:Cell Line; D018920:Coculture Techniques; D003594:Cytoplasmic Granules / Q000276:immunology; D003602:Cytotoxicity, Immunologic / Q000235:genetics*; D005786:Gene Expression Regulation / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D054992:Immunological Synapses; D007694:Killer Cells, Natural / Q000166:cytology / Q000276:immunology*; D008213:Lymphocyte Activation; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000235:genetics / Q000276:immunology*; D008840:Microfilament Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000276:immunology; D009154:Mutation; D010766:Phosphorylation; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014443:Tyrosine / Q000276:immunology / Q000378:metabolism; D051316:Wiskott-Aldrich Syndrome Protein / Q000235:genetics / Q000276:immunology
chemical_list: D008840:Microfilament Proteins; D034741:RNA, Small Interfering; C495767:WAS protein, human; C549919:WASH protein, human; D051316:Wiskott-Aldrich Syndrome Protein; D014443:Tyrosine; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck)