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Blood, Vol. 93 No. 4 (February 15), 1999:
pp. 1189-1196
Inhibitory Effect of the Transcription Factor Encoded by the
mi Mutant Allele in Cultured Mast Cells of Mice
Akihiko Ito,
Eiichi Morii,
Dae-Ki Kim,
Tatsuki R. Kataoka,
Tomoko Jippo,
Kazutaka Maeyama,
Hiroshi Nojima, and
Yukihiko Kitamura
From the Department of Pathology, Osaka University Medical School,
Suita, Osaka, Japan; the Department of Pharmacology, Ehime University
Medical School, Ehime, Japan; and the Department of Molecular Genetics,
Research Institute for Microbial Diseases, Osaka University, Suita,
Osaka, Japan.
The mi locus of mice encodes a transcription factor of the
basic-helix-loop-helix-leucine zipper protein family (MITF). The MITF
encoded by the mutant mi allele (mi-MITF) deletes 1 of
4 consecutive arginines in the basic domain. The mice of mi/mi
genotype express mi-MITF, whereas the mice of tg/tg
genotype have a transgene at the 5' flanking region of the
mi gene and do not express any MITF. To investigate the
function of mi-MITF in cultured mast cells (CMCs), we took two
approaches. First, mRNA obtained from mi/mi CMCs or
tg/tg CMCs was subtracted from complementary (c) DNA library of
normal (+/+) CMCs, and the (+/+-mi/mi) and
(+/+-tg/tg) subtraction libraries were obtained. When the
number of clones that hybridized more efficiently with +/+ CMC cDNA
probe than with mi/mi or tg/tg CMC cDNA probe was
compared using Southern analysis, the number was larger in the
(+/+-mi/mi) library than in the (+/+-tg/tg)
library. Second, we compared mRNA expression of six genes between
mi/mi and tg/tg CMCs by Northern analysis. The
transcription of three genes encoding mouse mast cell proteases was
impaired in both mi/mi and tg/tg CMCs. On the other
hand, the transcription of three genes encoding c-kit receptor,
tryptophan hydroxylase, and granzyme B was markedly reduced in
mi/mi CMCs, but the reduction was significantly smaller in
tg/tg CMCs. These results indicated the inhibitory effect of
mi-MITF on the transactivation of particular genes in CMCs.
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