Basal and antigen-induced exposure of the proline-rich sequence in CD3ε

Javier De La Cruz, Travis Kruger, Christopher A. Parks, Robert L. Silge, Nicolai S.C. Van Oers, Immanuel F. Luescher, Adam G. Schrum, Diana Gil

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15 Scopus citations

Abstract

The CD3ε cytoplasmic tail contains a conserved proline-rich sequence (PRS) that influences TCR-CD3 expression and signaling. Although the PRS can bind the SH3.1 domain of the cytosolic adapter Nck, whether the PRS is constitutively available for Nck binding or instead represents a cryptic motif that is exposed via conformational change upon TCR-CD3 engagement (CD3Δc) is currently unresolved. Furthermore, the extent to which a cis-acting CD3ε basic amino acid-rich stretch (BRS), with its unique phosphoinositide-binding capability, might impact PRS accessibility is not clear. In this study, we found that freshly harvested primary thymocytes expressed low to moderate basal levels of Nck-accessible PRS ("open-CD3"), although most TCR-CD3 complexes were inaccessible to Nck ("closed-CD3"). Ag presentation in vivo induced open-CD3, accounting for half of the basal level found in thymocytes from MHC+ mice. Additional stimulation with either anti-CD3 Abs or peptide-MHC ligands further elevated open-CD3 above basal levels, consistent with a model wherein antigenic engagement induces maximum PRS exposure. We also found that the open-CD3 conformation induced by APCs outlasted the time of ligand occupancy, marking receptors that had been engaged. Finally, CD3ε BRS-phosphoinositide interactions played no role in either adoption of the initial closed-CD3 conformation or induction of open-CD3 by Ab stimulation. Thus, a basal level of open-CD3 is succeeded by a higher, induced level upon TCR-CD3 engagement, involving CD3Δc and prolonged accessibility of the CD3ε PRS to Nck.

Original languageEnglish (US)
Pages (from-to)2282-2290
Number of pages9
JournalJournal of Immunology
Volume186
Issue number4
DOIs
StatePublished - Feb 15 2011

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ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

De La Cruz, J., Kruger, T., Parks, C. A., Silge, R. L., Van Oers, N. S. C., Luescher, I. F., Schrum, A. G., & Gil, D. (2011). Basal and antigen-induced exposure of the proline-rich sequence in CD3ε. Journal of Immunology, 186(4), 2282-2290. https://doi.org/10.4049/jimmunol.1003225