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

J Immunol. 2013 Apr 1;190(7):3749-56. doi: 10.4049/jimmunol.1203389. Epub 2013 Mar 4.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • Adaptor Proteins, Signal Transducing
  • Cell Adhesion Molecules
  • GRB2 Adaptor Protein
  • Intracellular Signaling Peptides and Proteins
  • LAT protein, human
  • Membrane Proteins
  • Nectins
  • thymocyte-expressed molecule involved in selection, human
  • ZAP-70 Protein-Tyrosine Kinase