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Blood, 15 January 2002, Vol. 99, No. 2, pp. 576-583
IMMUNOBIOLOGY
Cooperative binding of c-Myb and Pax-5 activates the
RAG-2 promoter in immature B cells
Hiroyuki Kishi,
Zhe-Xiong Jin,
Xing-Cheng Wei,
Takuya Nagata,
Tadashi Matsuda,
Shigeru Saito, and
Atsushi Muraguchi
From the Department of Immunology and the Department of
Obstetrics and Gynecology, Faculty of Medicine, Toyama Medical and
Pharmaceutical University, Sugitani, Toyama, Japan.
The recombination activating gene-1 (RAG-1) and
RAG-2 are expressed specifically in immature lymphoid cells
undergoing the recombination of antigen receptor genes. The regulation
of murine RAG-2 promoter was studied and it was revealed
that the  41/17 RAG-2 promoter region, which is
conserved between humans and mice, was indispensable for the
RAG-2 promoter activity in B-cell lines. The region
contained 2 cis elements that bound c-Myb and Pax-5. Mutation in the c-Myb-binding site in the promoter reduced the promoter activity in B-cell lines. Cooperative activation of the RAG-2 promoter was seen by a combination of c-Myb and Pax-5
in a human embryonic kidney cell line (293T), via their synergistic DNA-binding. Deletion experiments showed that the C-terminus of c-Myb
was responsible for their interaction. Furthermore, the dominant-negative c-Myb mutant suppressed the activation of the RAG-2 promoter in a pre-B-cell line as well as in 293T
cells. These results suggest that cooperative binding of c-Myb and
Pax-5 to the RAG-2 promoter is one of the mechanisms to
direct the restricted expression of the RAG-2 in immature B cells.
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