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Systematic Method to Obtain Novel Genes That Are Regulated by
mi Transcription Factor: Impaired Expression of Granzyme B and
Tryptophan Hydroxylase in mi/mi Cultured Mast Cells
Akihiko Ito,
Eiichi Morii,
Kazutaka Maeyama,
Tomoko Jippo,
Dae-Ki Kim,
Young-Mi Lee,
Hideki Ogihara,
Koji Hashimoto,
Yukihiko Kitamura, and
Hiroshi Nojima
From the Department of Pathology, Medical School, Osaka University,
Suita, Osaka; the Department of Pharmacology, Ehime University Medical
School, Ehime; and the Department of Molecular Genetics, Research
Institute for Microbial Diseases, Osaka University, Suita, Osaka,
Japan.
The mi locus encodes a member of the
basic-helix-loop-helix-leucine zipper protein family of transcription
factors (hereafter called MITF). We have reported that the expression
of several genes was impaired in cultured mast cells (CMCs) of
mi/mi genotype, and demonstrated the involvement of MITF in the
transcription of these genes. To obtain new genes whose transcription
may be regulated by MITF, we prepared a subtracted cDNA library using +/+ and mi/mi CMCs. We found two clones carrying the
granzyme (Gr) B and tryptophan hydroxylase (TPH) cDNAs in the
subtracted library. The expression of the Gr B and TPH genes decreased
in mi/mi CMCs, and recovered to nearly normal level by the
overexpression of normal (+) MITF but not of mutant (mi)
MITF. The +-MITF bound three and one CANNTG motifs in the Gr B and
TPH promoters, respectively, and transactivated these two genes,
indicating the involvement of +-MITF in their expression. Because TPH
is the rate-limiting enzyme for serotonin synthesis, we examined the
serotonin content of +/+ and mi/mi CMCs. The serotonin
content was significantly smaller in mi/mi CMCs than in +/+
CMCs. The introduction of +-MITF but not of mi-MITF
normalized the serotonin content in mi/mi CMCs.
Blood, Vol. 91 No. 9 (May 1), 1998:
pp. 3210-3221
© 1998 by The American Society of Hematology.
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