title: Dynein Separately Partners with NDE1 and Dynactin To Orchestrate T Cell Focused Secretion.
abstract: Helper and cytotoxic T cells accomplish focused secretion through the movement of vesicles toward the microtubule organizing center (MTOC) and translocation of the MTOC to the target contact site. In this study, using Jurkat cells and OT-I TCR transgenic primary murine CTLs, we show that the dynein-binding proteins nuclear distribution E homolog 1 (NDE1) and dynactin (as represented by p150(Glued)) form mutually exclusive complexes with dynein, exhibit nonoverlapping distributions in target-stimulated cells, and mediate different transport events. When Jurkat cells expressing a dominant negative form of NDE1 (NDE1-enhanced GFP fusion) were activated by Staphylococcus enterotoxin E-coated Raji cells, NDE1 and dynein failed to accumulate at the immunological synapse (IS) and MTOC translocation was inhibited. Knockdown of NDE1 in Jurkat cells or primary mouse CTLs also inhibited MTOC translocation and CTL-mediated killing. In contrast to NDE1, knockdown of p150(Glued), which depleted the alternative dynein/dynactin complex, resulted in impaired accumulation of CTLA4 and granzyme B-containing intracellular vesicles at the IS, whereas MTOC translocation was not affected. Depletion of p150(Glued) in CTLs also inhibited CTL-mediated lysis. We conclude that the NDE1/Lissencephaly 1 and dynactin complexes separately mediate two key components of T cell-focused secretion, namely translocation of the MTOC and lytic granules to the IS, respectively.
authors.affiliation: Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712; and.
authors.forename: Shubhankar
authors.identifier: 0000-0001-5778-7615
authors.initials: S
authors.lastname: Nath
authors.affiliation: Department of Molecular Cell and Developmental Biology, University of Texas at Austin, Austin, TX 78712.
authors.forename: Laura
authors.identifier: 0000-0003-3141-1710
authors.initials: L
authors.lastname: Christian
authors.affiliation: Department of Molecular Cell and Developmental Biology, University of Texas at Austin, Austin, TX 78712.
authors.forename: Sarah Youngsun
authors.identifier: 0000-0002-3428-4676
authors.initials: SY
authors.lastname: Tan
authors.affiliation: Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712; and.
authors.forename: Sanghee
authors.initials: S
authors.lastname: Ki
authors.affiliation: Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712; and.
authors.forename: Lauren I R
authors.identifier: 0000-0002-1697-1755
authors.initials: LI
authors.lastname: Ehrlich
authors.affiliation: Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712; and poenie@austin.utexas.edu.
authors.forename: Martin
authors.identifier: 0000-0003-2524-4039
authors.initials: M
authors.lastname: Poenie
mesh_terms: D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase / Q000502:physiology; D020013:Calcium Signaling; D003602:Cytotoxicity, Immunologic; D000072159:Dynactin Complex / Q000502:physiology*; D004398:Dyneins / Q000502:physiology*; D006801:Humans; D019169:Jurkat Cells; D008869:Microtubule-Associated Proteins / Q000502:physiology*; D022101:Microtubule-Organizing Center / Q000378:metabolism; D022142:Secretory Vesicles / Q000502:physiology; D013569:Synapses / Q000378:metabolism; D013601:T-Lymphocytes / Q000502:physiology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins; C525094:Nde1 protein, human; D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase; C084102:PAFAH1B1 protein, human; D004398:Dyneins