Evidence for inducible recruitment of Wiskott-Aldrich syndrome protein to T cell receptor-CD3 complex in Jurkat T cells.
BACKGROUND
The engagement of the T cell receptor (TCR)-CD3 complex induces the formation of multiple signalling complexes, which are required for actin cytoskeletal rearrangement. The Wiskott-Aldrich syndrome protein (WASp) is a key regulator of actin polymerization that is recruited to the TCR activation site. Since WASp is a binding partner of adaptor protein Nck, which is recruited directly to the TCR CD3? subunit upon TCR ligation, therefore we proposed that the direct recruitment of Nck to TCR-CD3 may also bring WASp directly to TCR-CD3.


OBJECTIVE
The aim of this present study was to assess the distribution of WASp, in relation to Nck, to the TCR-CD3ε complex.


METHODS
Jurkat T cells were stimulated with anti-TCR antibody and then the cell lysates were immunoprecipitated with anti-CD3 antibody before immunoblotting with antibodies specific to WASp, Nck1, Nck2, SLP-76 and CD3ε molecules.


RESULTS
WASp was recruited to SLP-76 and also directly to the TCR-CD3 complex upon TCR triggering. The inducible recruitment of WASp to the TCR-CD3 complex is partially dependent of tyrosine phosphorylation.


CONCLUSIONS
The present findings provide an alternative mechanism of WASp recruitment to the site of TCR activation that may be involved in recruitment of Nck.
Department of Microbiology and Parasitology, Faculty of Medical Science; Naresuan University, Pitsanulok 65000 Thailand.
Pussadee
P
Paensuwan
Jatuporn
J
Ngoenkam
Boonruang
B
Khamsri
Kanlaya
K
Preechanukul
Donruedee
D
Sanguansermsri
Sutatip
S
Pongcharoen
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D017252:CD3 Complex / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D008213:Lymphocyte Activation; D015513:Oncogene Proteins / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D021381:Protein Transport; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000276:immunology / Q000378:metabolism*; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D014443:Tyrosine; D051316:Wiskott-Aldrich Syndrome Protein / Q000276:immunology / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D017252:CD3 Complex; C115335:CD3E protein, human; C078332:Nck protein; D015513:Oncogene Proteins; D010750:Phosphoproteins; D017260:Receptor-CD3 Complex, Antigen, T-Cell; C092947:SLP-76 signal Transducing adaptor proteins; C495767:WAS protein, human; D051316:Wiskott-Aldrich Syndrome Protein; D014443:Tyrosine