TY - JOUR
T1 - Intracellular Redistribution of Nucleolin upon Interaction with the CD3ε Chain of the T Cell Receptor Complex
AU - Gil, Diana
AU - Gutiérrez, Dolores
AU - Alarcón, Balbino
PY - 2001/4/6
Y1 - 2001/4/6
N2 - 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.
AB - 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.
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U2 - 10.1074/jbc.M010114200
DO - 10.1074/jbc.M010114200
M3 - Article
C2 - 11115514
AN - SCOPUS:0035815623
SN - 0021-9258
VL - 276
SP - 11174
EP - 11179
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 14
ER -