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Mechanistic basis of pre–T cell receptor–mediated autonomous signaling critical for thymocyte development

Nature Immunology volume 7, pages 67–75 (2006)Cite this article

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Abstract

The pre–T cell receptor (TCR) is crucial for early T cell development and is proposed to function in a ligand-independent way. However, the molecular mechanism underlying the autonomous signals remains elusive. Here we show that the pre-TCR complex spontaneously formed oligomers. Specific charged residues in the extracellular domain of the pre-TCR α-chain mediated formation of the oligomers in vitro. Alteration of these residues eliminated the ability of the pre-TCR α-chain to support pre-TCR signaling in vivo. Dimerization but not raft localization of CD3ε was sufficient to simulate pre-TCR function and promote β-selection. These results suggest that the pre-TCR complex can deliver its signal autonomously through oligomerization of the pre-TCR α-chain mediated by charged residues.

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Figure 1: Constitutive internalization and lysosome targeting of the pre-TCR.
Figure 2: Autonomous growth of BAF3 cell lines is induced by pTα–EPOR but not TCRα–EPOR.
Figure 3: Specific charged amino acids in the extracellular domain of pTα are critical for the function of pTα–EPOR.
Figure 4: The pTα-RA mutant fails to induce pre-TCR oligomerization or to restore Ptcra−/− thymocyte development.
Figure 5: Raft localization of CD3ε is not sufficient for the DP transition.
Figure 6: Dimerized CD3ε chimera is sufficient for bypass of β-selection in Rag2−/− mice.
Figure 7: The CD3ε dimer induces differentiation signals in DN cells but not DP cells or a mature T cell line.

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Protein Data Bank

Change history

  • 09 December 2005

    In the version of this article initially published online, the fifth subheading in the Results section was incorrect. The error has been corrected for the HTML and print versions of the article.

Notes

  1. *Note: In the version of this article initially published online, the fifth subheading in the Results section was incorrect. It should read “Raft-targeted CD3ε fails to bypass β-selection.” The error has been corrected for the HTML and print versions of the article.

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Acknowledgements

We thank A. Yoshimura for cDNA; M. Unno and R. Shiina for technical help; H. Tomiyama for FRET analysis; N. Shinohara, S. Koyasu, A. Kosugi, H. Suzuki, K. Masuda, S. Sugawa, H. Fukuyama and O. Kanagawa for discussions; H. von Boehmer and S. Taki for critical reading of the manuscript; and H. Yamaguchi for secretarial assistance. Supported by a Grant-in-Aid for Priority Area Research (A) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Authors and Affiliations

  1. Laboratory for Cell Signaling, RIKEN Research Center for Allergy and Immunology, Yokohama, 230-0045, Kanagawa, Japan

    Sho Yamasaki, Eri Ishikawa, Machie Sakuma & Takashi Saito

  2. Centre for Computational Biology, The Hospital for Sick Children, Toronto, M5G 1X8, Ontario, Canada

    Koji Ogata

  3. Single Molecule Immunoimaging, RIKEN Research Center for Allergy and Immunology, Yokohama, 230-0045, Kanagawa, Japan

    Kumiko Sakata-Sogawa, Michio Hiroshima & Makio Tokunaga

  4. Fox Chase Cancer Center, Philadelphia, 19111, Pennsylvania, USA

    David L Wiest

  5. Structural Biology Center, National Institute of Genetics, and Department of Genetics, The Graduate University for Advanced Studies, Shizuoka, 411-8540, Japan

    Makio Tokunaga

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Supplementary information

Supplementary Fig. 1

pre-TCR has the tendency to form oligomers on the cell surface. (PDF 11 kb)

Supplementary Fig. 2

Generation of FRET in T cells co-expressing pTα–CFP and pTα–YFP. (PDF 45 kb)

Supplementary Fig. 3

Competitive BMT reconsitution assay of pTα-WT and pTα-R24A. (PDF 10 kb)

Supplementary Fig. 4

hCD8–CD3e chimera induced DN3 to DN4 cell transition in the absence of pTα. (PDF 29 kb)

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Yamasaki, S., Ishikawa, E., Sakuma, M. et al. Mechanistic basis of pre–T cell receptor–mediated autonomous signaling critical for thymocyte development. Nat Immunol 7, 67–75 (2006). https://doi.org/10.1038/ni1290

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