title: The adaptor protein GULP promotes Jedi-1-mediated phagocytosis through a clathrin-dependent mechanism.
abstract: During the development of the peripheral nervous system, the large number of apoptotic neurons generated are phagocytosed by glial precursor cells. This clearance is mediated, in part, through the mammalian engulfment receptor Jedi-1. However, the mechanisms by which Jedi-1 mediates phagocytosis are poorly understood. Here we demonstrate that Jedi-1 associates with GULP, the mammalian homologue of CED-6, an adaptor protein required for phagocytosis mediated by the nematode engulfment receptor CED-1. Silencing GULP or mutating the NPXY motif in Jedi-1, which is required for GULP binding, prevents Jedi-1-mediated phagocytosis. How GULP promotes engulfment is not known. Of interest, we find that Jedi-1-induced phagocytosis requires GULP binding to clathrin heavy chain (CHC). During engulfment, CHC is tyrosine phosphorylated, which is required for Jedi-mediated engulfment. Both phosphoclathrin and actin accumulate around engulfed microspheres. Furthermore, knockdown of CHC in HeLa cells prevents Jedi-1-mediated engulfment of microspheres, and knockdown in glial precursors prevents the engulfment of apoptotic neurons. Taken together, these results reveal that Jedi-1 signals through recruitment of GULP, which promotes phagocytosis through a noncanonical phosphoclathrin-dependent mechanism.
authors.affiliation: Department of Biochemistry, Vanderbilt Brain Institute, Vanderbilt University School of Medicine, Nashville, TN 37232.
authors.forename: Chelsea S
authors.initials: CS
authors.lastname: Sullivan
authors.affiliation: Department of Biochemistry, Vanderbilt Brain Institute, Vanderbilt University School of Medicine, Nashville, TN 37232.
authors.forename: Jami L
authors.initials: JL
authors.lastname: Scheib
authors.affiliation: Center for Cell Clearance and Department of Microbiology, Immunology, and Cancer Biology, University of Virginia School of Medicine, Charlottesville, VA 22908.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Ma
authors.affiliation: Department of Biochemistry, Vanderbilt Brain Institute, Vanderbilt University School of Medicine, Nashville, TN 37232.
authors.forename: Rajan P
authors.initials: RP
authors.lastname: Dang
authors.affiliation: Department of Biochemistry, Vanderbilt Brain Institute, Vanderbilt University School of Medicine, Nashville, TN 37232.
authors.forename: Johanna M
authors.initials: JM
authors.lastname: Schafer
authors.affiliation: Department of Biochemistry, Vanderbilt Brain Institute, Vanderbilt University School of Medicine, Nashville, TN 37232.
authors.forename: Francis E
authors.initials: FE
authors.lastname: Hickman
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, Department of Microbiology and Immunology, and Department of Pharmaceutical Chemistry, G. W. Hooper Foundation, University of California, San Francisco, San Francisco, CA 94143.
authors.forename: Frances M
authors.initials: FM
authors.lastname: Brodsky
authors.affiliation: Center for Cell Clearance and Department of Microbiology, Immunology, and Cancer Biology, University of Virginia School of Medicine, Charlottesville, VA 22908.
authors.forename: Kodi S
authors.initials: KS
authors.lastname: Ravichandran
authors.affiliation: Department of Biochemistry, Vanderbilt Brain Institute, Vanderbilt University School of Medicine, Nashville, TN 37232 bruce.carter@vanderbilt.edu.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Carter
mesh_terms: D016475:3T3 Cells; D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000502:physiology*; D020816:Amino Acid Motifs; D000818:Animals; D002966:Clathrin / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D010587:Phagocytosis; D054730:Protein Interaction Domains and Motifs; D021381:Protein Transport
chemical_list: D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D002966:Clathrin; C000594793:GULP protein, mouse; C522125:Jedi protein, mouse; D008565:Membrane Proteins
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