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and Dad1 Genes1
Department of Immunology, Duke University Medical Center, Durham, NC 27710
Although tightly linked, the TCR 
and 
genes are expressed
specifically in T lymphocytes, whereas the Dad1 gene is ubiquitously
expressed. Between TCR and Dad1 are eight DNase I hypersensitive
sites (HS). HS1 colocalizes with the TCR enhancer
(E) and is T cell-specific; HS2, -3, -4, -5, and -6 map
downstream of HS1 and are tissue-nonspecific. The region spanning
HS2–6 was reported to display chromatin-opening activity and to confer
copy number-dependent and integration site-independent transgene
expression in transgenic mice. Here, we demonstrate that HS2–6 also
displays enhancer-blocking activity, as it can block an enhancer from
activating a promoter when located between the two in a
chromatin-integrated context, and can do so without repressing either
the enhancer or the promoter. Multiple enhancer-blocking elements are
arrayed across HS2–6. We show that HS2–6 by itself does not activate
transcription in chromatin context, but can synergize with an enhancer
when located upstream of an enhancer and promoter. We propose that
HS2–6 primarily functions as an insulator or boundary element that may
be critical for the autonomous regulation of the TCR and Dad1
genes.
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