The Journal of Experimental Medicine
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Published online 1 October 2001. doi:10.1084/jem.194.7.979
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© The Rockefeller University Press, 0022-1007/2001/10/979/ $5.00
The Journal of Experimental Medicine, Volume 194, Number 7, October 1, 2001 979-990

Original Article

The Upstream Enhancer Is Necessary and Sufficient for the Expression of the Pre-T Cell Receptor {alpha} Gene in Immature T Lymphocytes

Boris Reizisa and Philip Ledera

a Department of Genetics and Howard Hughes Medical Institute, Harvard Medical School, Boston, MA 02115
Dept. of Genetics, Harvard Medical School, 200 Longwood Ave., Boston, MA 02115.617-432 7944617-432-7667

leder{at}rascal.med.harvard.edu

The expression of the pre-T cell receptor {alpha} (pTa) gene occurs exclusively in immature T lymphocytes and is regulated by poorly defined mechanisms. We have analyzed the role of the upstream enhancer in pTa expression using conventional and bacterial artificial chromosome (BAC) reporter transgenes. The deletion of the enhancer completely abolished the expression of pTa BAC reporter in transgenic mice. Conversely, the combination of pTa enhancer and promoter targeted transgenes specifically to immature thymocytes, recapitulating the expression pattern of pTa. The core enhancer is conserved between mice and humans and contains a critical binding site for the transcription factor c-Myb. We also show that pTa promoter contains a conserved tandem E box site activated by E protein, HEB. These data establish the enhancer as a critical element regulating pTa gene expression and identify additional targets for c-Myb and E proteins in T cell development.

Key Words: transcription • thymus • c-Myb • basic helix-loop-helix proteins • HEB

Abbreviations used in this paper: APC, allophycocyanin; BAC, bacterial artificial chromosome; bHLH, basic helix-loop-helix; DBD, DNA-binding domain; DN, double-negative; DP, double-positive; EGFP, enhanced green fluorescent protein; EMSA, electrophoretic mobility shift assay; ISP, immature single-positive; pTa, pre-T cell receptor {alpha}; SP, single-positive.

© 2001 The Rockefeller University Press


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