Intracellular Redistribution of Nucleolin upon Interaction with the CD3ε Chain of the T Cell Receptor Complex

Diana Gil, Dolores Gutiérrez, Balbino Alarcón

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

T cell activation through the antigen receptor (TCR) involves the cytoplasmic tails of the CD3 subunits CD3γ, CD3δ, CD3ε, and CD3ζ. Whereas the biological significance of the cytoplasmic tails of these molecules is suggested, in part, by their evolutionarily conserved sequences, their interactions with signal transduction molecules are not completely understood. We used affinity chromatography columns of glutathione S-transferase fused to the CD3ε cytoplasmic tail to isolate proteins that specifically interact with this subunit. In this way, we identified the shuttling protein nucleolin as a specific CD3ε-interacting molecule. Using competition studies and affinity chromatography on peptide columns, we were able to identify a central proline-rich sequence as the nucleolin-interacting sequence in CD3ε. Transfection in COS cells of wild type CD3ε, but not of nonbinding mutants of CD3ε, resulted in redistribution of nucleolin from the nucleus and nucleoli to the cytoplasm. This property was transferred to a CD8 protein chimera by appending the cytoplasmic tail of CD3ε. We also found that nucleolin associated with the TCR complex. This association was increased upon TCR engagement, suggesting that the CD3ε/nucleolin interaction may have a role in T cell activation.

Original languageEnglish (US)
Pages (from-to)11174-11179
Number of pages6
JournalJournal of Biological Chemistry
Volume276
Issue number14
DOIs
StatePublished - Apr 6 2001

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Intracellular Redistribution of Nucleolin upon Interaction with the CD3ε Chain of the T Cell Receptor Complex'. Together they form a unique fingerprint.

Cite this