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Chromatin structure and transcriptional regulation of human RAG-1 gene
T Kitagawa, K Mori, H Kishi, H Tagoh, T Nagata, H Kurioka and A Muraguchi
Department of Immunology, Faculty of Medicine, Toyama Medical and
Pharmaceutical University, Japan.
The recombination activating genes (RAGs) play a critical role in V(D)J
recombination machinery and lymphocyte development. Their expression is
strictly regulated during lymphocyte ontogeny, with expression being
rapidly lost as the lymphoid precursors differentiate into their progeny.
To elucidate molecular mechanisms of regulation of human RAG-1 gene
expression, we examined a chromatin structure of a approximately 24-kb DNA
segment adjacent to a human RAG-1 promoter region in various cell lines by
analyzing DNase I hypersensitive (DNase I HS) sites. In a RAG-1-expressing
human pre-B-cell line, at least four DNase I HS sites (HS1, HS2, HS3, and
HS4) were identified. Among these HS sites, one HS site (HS1) was
ubiquitously detected in all cell lines examined, but the other three HS
sites (HS2, HS3, and HS4) were associated only with RAG-1-expressing
lymphoid cell lines. Using transient expression assays, we showed that the
5' upstream region of the major transcription start site showed low but
significant promoter activity and that a DNA segment within HS3 located in
the promoter region was indispensable to its basal promoter activity.
Importantly, this promoter region was shown to be active in both
RAG-1-expressing and RAG- 1-nonexpressing cell lines. These results suggest
that alteration of chromatin structure in the promoter region, in addition
to other control elements outside of the promoter region, is one of the
mechanisms regulating tissue- and stage-specific expression of human RAG-1
gene.
Volume 88,
Issue 10,
pp. 3785-3791,
11/15/1996
Copyright © 1996 by The American Society of Hematology
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