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Functional Analysis of the Murine TCRß-Chain Gene Enhancer¹

Rosenstiel Research Center and Department of Biology, Brandeis University, Waltham, MA 02254

The TCRß-chain gene enhancer activates transcription and V(D)J recombination in immature thymocytes. In this paper we present a systematic analysis of the elements that contribute to the activity of the murine TCRß enhancer in mature and immature T cell lines. We identified a region containing the ßE4, ßE5, and ßE6 motifs as the essential core of the TCRß enhancer in pro-T cells. In mature cells, the core enhancer had low activity and required, in addition, either 5' or 3' flanking sequences whose functions may be partially overlapping. Mutation of any of the six protein binding sites located within the ßE4–ßE6 elements essentially abolished enhancer activity, indicating that this core enhancer contained no redundant elements. The ßE4 and ßE6 elements contain binding sites for ETS-domain proteins and the core binding factor. The ßE5 element bound two proteins that could be resolved chromatographically and that were both essential for enhancer activity.