title: T-cell receptor ligation causes Wiskott-Aldrich syndrome protein degradation and F-actin assembly downregulation.
abstract: BACKGROUND
  Wiskott-Aldrich syndrome protein (WASP) links T-cell receptor (TCR) signaling to the actin cytoskeleton. WASP is normally protected from degradation by the Ca(++)-dependent protease calpain and by the proteasome because of its interaction with the WASP-interacting protein.
  
  
  OBJECTIVE
  We investigated whether WASP is degraded after TCR ligation and whether its degradation downregulates F-actin assembly caused by TCR ligation.
  
  
  METHODS
  Primary T cells, Jurkat T cells, and transfected 293T cells were used in immunoprecipitation experiments. Intracellular F-actin content was measured in splenic T cells from wild-type, WASP-deficient, and c-Casitas B-lineage lymphoma (Cbl)-b-deficient mice by using flow cytometry. Calpeptin and MG-132 were used to inhibit calpain and the proteasome, respectively.
  
  
  RESULTS
  A fraction of WASP in T cells was degraded by calpain and by the ubiquitin-proteasome pathway after TCR ligation. The Cbl-b and c-Cbl E3 ubiquitin ligases associated with WASP after TCR signaling and caused its ubiquitination. Inhibition of calpain and lack of Cbl-b resulted in a significantly more sustained increase in F-actin content after TCR ligation in wild-type T cells but not in WASP-deficient T cells.
  
  
  CONCLUSIONS
  TCR ligation causes WASP to be degraded by calpain and to be ubiquitinated by Cbl family E3 ligases, which targets it for destruction by the proteasome. WASP degradation might provide a mechanism for regulating WASP-dependent TCR-driven assembly of F-actin.
authors.affiliation: Department of Pediatrics, Tohoku University Graduate School of Medicine, Miyagi, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Watanabe
authors.affiliation: Department of Pediatrics, Tohoku University Graduate School of Medicine, Miyagi, Japan. Electronic address: ysasahara@med.tohoku.ac.jp.
authors.forename: Yoji
authors.initials: Y
authors.lastname: Sasahara
authors.affiliation: Division of Immunology, Children's Hospital, and the Department of Pediatrics, Harvard Medical School, Boston, Mass.
authors.forename: Narayanaswamy
authors.initials: N
authors.lastname: Ramesh
authors.affiliation: Division of Immunology, Children's Hospital, and the Department of Pediatrics, Harvard Medical School, Boston, Mass.
authors.forename: Michel J
authors.initials: MJ
authors.lastname: Massaad
authors.affiliation: Department of Pediatrics, Tohoku University Graduate School of Medicine, Miyagi, Japan.
authors.forename: Chung
authors.initials: C
authors.lastname: Yeng Looi
authors.affiliation: Department of Pediatrics, Tohoku University Graduate School of Medicine, Miyagi, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Kumaki
authors.affiliation: Department of Pediatrics, Tohoku University Graduate School of Medicine, Miyagi, Japan.
authors.forename: Shigeo
authors.initials: S
authors.lastname: Kure
authors.affiliation: Division of Immunology, Children's Hospital, and the Department of Pediatrics, Harvard Medical School, Boston, Mass.
authors.forename: Raif S
authors.initials: RS
authors.lastname: Geha
authors.affiliation: Department of Pediatrics, Tohoku University Graduate School of Medicine, Miyagi, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Tsuchiya
mesh_terms: D000199:Actins / Q000378:metabolism*; D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000818:Animals; D002154:Calpain / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004151:Dipeptides / Q000494:pharmacology; D015536:Down-Regulation; D007976:Leupeptins / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D050721:Proto-Oncogene Proteins c-cbl / Q000378:metabolism*; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism; D054875:Ubiquitination; D051316:Wiskott-Aldrich Syndrome Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; C118387:Cblb protein, mouse; D015853:Cysteine Proteinase Inhibitors; D004151:Dipeptides; D007976:Leupeptins; D011948:Receptors, Antigen, T-Cell; D051316:Wiskott-Aldrich Syndrome Protein; C071482:calpeptin; D050721:Proto-Oncogene Proteins c-cbl; D002154:Calpain; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde
keywords: Actin-related protein; Arp; Casitas B-lineage lymphoma; Cbl; Cbl family proteins; EVH1; Ena-VASP homology domain 1; F-actin; IS; Immune synapse; T-cell receptor; TCR; WAS; WASP; WASP-interacting protein; WIP; WT; Wild-type; Wiskott-Aldrich syndrome; Wiskott-Aldrich syndrome protein; ZAP-70; calpain; proteasome; ubiquitination; ζ Chain–associated protein kinase of 70 kDa