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Regulation of mouse mast cell protease 6 gene expression by transcription
factor encoded by the mi locus
E Morii, T Tsujimura, T Jippo, K Hashimoto, K Takebayashi, K Tsujino, S Nomura, M Yamamoto and Y Kitamura
Department of Pathology, Osaka University Medical School, Japan.
The mi locus of mice encodes a member of the basic-helix-loop-helix-
leucine zipper (bHLH-Zip) protein family of transcription factors
(hereafter called MITF). Because the expression of the mouse mast cell
protease 6 (MMCP-6) gene is remarkably reduced in mast cells of mi/mi
mutant mice, we investigated the effect of MITF on the transcription of the
MMCP-6 gene. First, we introduced the normal (+) MITF cDNA into mi/mi
cultured mast cells using the retroviral vector. Overexpression of +-MITF
but not mi-MITF normalized the expression of the MMCP-6 gene, indicating
the involvement of +-MITF in the MMCP-6 gene transactivation. Second, we
analyzed the promoter of the MMCP-6 gene by the transient cotransfection
assay. The luciferase construct under the control of the MMCP-6 promoter
and the cDNA encoding +-MITF or mi-MITF were cotransfected into NIH/ 3T3
fibroblasts. The coexpression of +- MITF but not mi-MITF increased the
luciferase activity 10-fold. We found a CACATG and a CATCTG motif in the
MMCP-6 promoter, both of which are generally recognized by bHLH-Zip-type
transcription factors. We also found a GACCTG motif that was strongly bound
by +-MITF. These three motifs were necessary for the 10-fold
transactivation ability of the MMCP-6 promoter by +-MITF. Mutations of each
motif significantly reduced the transactivation, suggesting that +-MITF
directly transactivated the MMCP-6 gene through these three motifs.
Volume 88,
Issue 7,
pp. 2488-2494,
10/01/1996
Copyright © 1996 by The American Society of Hematology
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