title: Myosin IIA associates with NK cell lytic granules to enable their interaction with F-actin and function at the immunological synapse.
abstract: NK cell cytotoxicity requires the formation of an actin-rich immunological synapse (IS) with a target cell and the polarization of perforin-containing lytic granules toward the IS. Following the polarization of lytic granules, they traverse through the actin-rich IS to join the NK cell membrane in order for directed secretion of their contents to occur. We examined the role of myosin IIA as a candidate for facilitating this prefinal step in lytic NK cell IS function. Lytic granules in and derived from a human NK cell line, or ex vivo human NK cells, were constitutively associated with myosin IIA. When isolated using density gradients, myosin IIA-associated NK cell lytic granules directly bound to F-actin and the interaction was sensitive to the presence of ATP under conditions of flow. In NK cells from patients with a truncation mutation in myosin IIA, NK cell cytotoxicity, lytic granule penetration into F-actin at the IS, and interaction of isolated granules with F-actin were all decreased. Similarly, inhibition of myosin function also diminished the penetration of lytic granules into F-actin at the IS, as well as the final approach of lytic granules to and their dynamics at the IS. Thus, NK cell lytic granule-associated myosin IIA enables their interaction with actin and final transit through the actin-rich IS to the synaptic membrane, and can be defective in the context of naturally occurring human myosin IIA mutation.
authors.affiliation: Immunology Graduate Group, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
authors.forename: Keri B
authors.initials: KB
authors.lastname: Sanborn
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Rak
authors.forename: Saumya Y
authors.initials: SY
authors.lastname: Maru
authors.forename: Korey
authors.initials: K
authors.lastname: Demers
authors.forename: Analisa
authors.initials: A
authors.lastname: Difeo
authors.forename: John A
authors.initials: JA
authors.lastname: Martignetti
authors.forename: Michael R
authors.initials: MR
authors.lastname: Betts
authors.forename: Rémi
authors.initials: R
authors.lastname: Favier
authors.forename: Pinaki P
authors.initials: PP
authors.lastname: Banerjee
authors.forename: Jordan S
authors.initials: JS
authors.lastname: Orange
mesh_terms: D000199:Actins / Q000378:metabolism*; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D003594:Cytoplasmic Granules / Q000378:metabolism*; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology* / Q000648:ultrastructure; D009154:Mutation; D024507:Nonmuscle Myosin Type IIA / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D024507:Nonmuscle Myosin Type IIA