GRB2-mediated recruitment of THEMIS to LAT is essential for thymocyte development

Wolfgang Paster, Claudia Brockmeyer, Guo Fu, Philip C Simister, Ben de Wet, Ana Martinez-Riaño, John A H Hoerter, Stephan M Feller, Christoph Wülfing, Nicholas R J Gascoigne, Oreste Acuto, Christoph Wülfing

Research output: Contribution to journalArticle (Academic Journal)

49 Citations (Scopus)

Abstract

Thymocyte-expressed molecule involved in selection (THEMIS) is a recently identified regulator of thymocyte positive selection. THEMIS's mechanism of action is unknown, and whether it has a role in TCR-proximal signaling is controversial. In this article, we show that THEMIS and the adapter molecule growth factor receptor-bound protein 2 (GRB2) associate constitutively through binding of a conserved PxRPxK motif within the proline-rich region 1 of THEMIS to the C-terminal SH3-domain of GRB2. This association is indispensable for THEMIS recruitment to the immunological synapse via the transmembrane adapter linker for activation of T cells (LAT) and for THEMIS phosphorylation by Lck and ZAP-70. Two major sites of tyrosine phosphorylation were mapped to a YY-motif close to proline-rich region 1. The YY-motif was crucial for GRB2 binding, suggesting that this region of THEMIS might control local phosphorylation-dependent conformational changes important for THEMIS function. Finally, THEMIS binding to GRB2 was required for thymocyte development. Our data firmly assign THEMIS to the TCR-proximal signaling cascade as a participant in the LAT signalosome and suggest that the THEMIS-GRB2 complex might be involved in shaping the nature of Ras signaling, thereby governing thymic selection.

Original languageEnglish
Pages (from-to)3749-56
Number of pages8
JournalJournal of Immunology
Volume190
Issue number7
DOIs
Publication statusPublished - 1 Apr 2013

Keywords

  • Adaptor Proteins, Signal Transducing
  • Amino Acid Sequence
  • Cell Adhesion Molecules
  • Cell Line
  • GRB2 Adaptor Protein
  • Humans
  • Immunological Synapses
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Molecular Sequence Data
  • Phosphorylation
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Sequence Alignment
  • Thymocytes
  • ZAP-70 Protein-Tyrosine Kinase

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    Paster, W., Brockmeyer, C., Fu, G., Simister, P. C., de Wet, B., Martinez-Riaño, A., Hoerter, J. A. H., Feller, S. M., Wülfing, C., Gascoigne, N. R. J., Acuto, O., & Wülfing, C. (2013). GRB2-mediated recruitment of THEMIS to LAT is essential for thymocyte development. Journal of Immunology, 190(7), 3749-56. https://doi.org/10.4049/jimmunol.1203389