title: Structure/function relationship of activating Ly-49D and inhibitory Ly-49G2 NK receptors.
abstract: Murine NK cells express Ly-49 family receptors capable of either inhibiting or activating lytic function. The overlapping patterns of expression of the various receptors have complicated their precise biochemical characterization. Here we describe the use of the Jurkat T cell line as the model for the study of Ly-49s. We demonstrate that Ly-49D is capable of delivering activation signals to Jurkat T cells even in the absence of the recently described Ly-49D-associated chain, DAP-12. Ly-49D signaling in Jurkat leads to tyrosine phosphorylation of TCRzeta and requires Syk/Zap70 family kinases and arginine 54 of Ly-49D, suggesting that Ly-49D signals via association with TCRzeta. Coexpression studies in 293-T cells confirmed the ability of Ly-49D to associate with TCRzeta. In addition, we have used this model to study the functional interactions between an inhibitory Ly-49 (Ly-49G2) and an activating Ly-49 (Ly-49D). Ly-49G2 blocks activation mediated by Ly-49D in an immunoreceptor tyrosine-based inhibitory motif (ITIM)-dependent manner. In contrast, Ly-49G2 was incapable of inhibiting activation by the TCR even though human killer cell inhibitory receptor (KIR) (KIR3DL2(GL183)) effectively inhibits TCR. Both the ability of Ly-49G2 to block Ly-49D activation and the failure of Ly-49G2 to inhibit TCR signaling were confirmed in primary murine NK cells and NK/T cells, respectively. These data demonstrate the dominant effects of the inhibitory receptors over those that activate and suggest an inability of the Ly-49 type II inhibitory receptors to efficiently inhibit type I transmembrane receptor signaling in T cells and NK cells.
authors.affiliation: Laboratory of Experimental Immunology, Division of Basic Sciences, Science Applications International Corporation-Frederick, MD 21702, USA. Ortaldo@mail.ncifcrf.gov
authors.forename: J R
authors.initials: JR
authors.lastname: Ortaldo
authors.forename: R
authors.initials: R
authors.lastname: Winkler-Pickett
authors.forename: J
authors.initials: J
authors.lastname: Willette-Brown
authors.forename: R L
authors.initials: RL
authors.lastname: Wange
authors.forename: S K
authors.initials: SK
authors.lastname: Anderson
authors.forename: G J
authors.initials: GJ
authors.lastname: Palumbo
authors.forename: L H
authors.initials: LH
authors.lastname: Mason
authors.forename: D W
authors.initials: DW
authors.lastname: McVicar
mesh_terms: D020816:Amino Acid Motifs / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000950:Antigens, Ly / Q000737:chemistry / Q000502:physiology; D020013:Calcium Signaling / Q000276:immunology; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D019169:Jurkat Cells; D007694:Killer Cells, Natural / Q000276:immunology / Q000378:metabolism*; D037181:Lectins, C-Type; D008213:Lymphocyte Activation; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000737:chemistry* / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000737:chemistry* / Q000502:physiology*; D054340:Receptors, KIR; D054347:Receptors, KIR3DL2; D055630:Receptors, NK Cell Lectin-Like; D015398:Signal Transduction / Q000276:immunology; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
chemical_list: D000950:Antigens, Ly; D007376:Interleukin-2; C515084:KIR3DL2 protein, human; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D011971:Receptors, Immunologic; D054340:Receptors, KIR; D054347:Receptors, KIR3DL2; D055630:Receptors, NK Cell Lectin-Like; D014443:Tyrosine