Dynamic molecular interactions linking the T cell antigen receptor to the actin cytoskeleton

Abstract

T cell receptor (TCR) engagement leads to actin polymerization at the site of T cell contact with antigen-presenting cells. Here we have studied the dynamic activity of proteins involved in regulating actin polymerization in live T cells after activation. Two such adaptor proteins, Nck and the Wiskott-Aldrich syndrome protein (WASp), were recruited to the TCR during initial T cell activation, where they colocalized with the tyrosine kinase Zap70. The recruitment of Nck and WASp depended on TCR-induced tyrosine phosphorylation and the LAT and SLP-76 adaptors. Nck and WASp migrated peripherally and accumulated at an actin-rich circumferential ring. Thus, actin polymerization regulated by the TCR begins at the TCR. Molecules recruited to the TCR regulate actin polymerization and this process drives plasma membrane movement and cellular spreading.

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Figure 1: WASp is recruited to the activated TCR site and then migrates to sites of actin rearrangement.
Figure 2: Characterization of the molecular interaction between SLP-76 and WASp in fixed and live cells.
Figure 3: The Nck molecule is dynamically redistributed throughout contact formation.
Figure 4: Nck recruitment to the T cell contact site is dependent on the functional activity of LAT.
Figure 5: Nck recruitment to the T cell contact site is dependent on SLP-76.

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Acknowledgements

The authors thank S. Saitoh and D. Mahadeo for their help; S. Garfield and S. Wincovitch for their help in the operation of the LSM 510; B. Taylor for cell sorting; and P. Schwartzberg and C. Sommers for reading the manuscript. Supported by the Cancer Research Institute (S.C.B.) and the Office of Research on Women's Health, Foundation for Advanced Education in the Sciences, National Institutes of Health (R.T.).

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Correspondence to Lawrence E Samelson.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Known and proposed molecular interactions leading to TCR-induced actin polymerization. (PDF 64 kb)

Supplementary Fig. 2

FRET analysis controls. (PDF 72 kb)

Supplementary Fig. 3

Reconstitution of Jurkat cells with the lipid phosphatase PTEN. (PDF 177 kb)

Supplementary Video 1

Dynamic localization of WASp-YFP actin-CFP in live activated T cells. (MOV 431 kb)

Supplementary Video 2

Dynamic localization of SLP-76-YFP WASp-CFP in live activated T cells. (MOV 735 kb)

Supplementary Video 3

Dynamic localization of SLP-76-YFP CFP-Nck in live activated T cells. (MOV 5311 kb)

Supplementary Video 4

Dynamic localization of YFP-Nck WASp-CFP in live activated T cells. (MOV 660 kb)

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Barda-Saad, M., Braiman, A., Titerence, R. et al. Dynamic molecular interactions linking the T cell antigen receptor to the actin cytoskeleton. Nat Immunol 6, 80–89 (2005). https://doi.org/10.1038/ni1143

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