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-Melanocyte-Stimulating Hormone Receptor in Cultured Mast Cells of Mice1 ,2
Department of Pathology, Osaka University Medical School, Suita, Japan
The microphthalmia (mi) locus encodes a member of
the basic-helix-loop-helix-leucine zipper (bHLH-Zip) protein family of
transcription factors (MITF). We have reported that expression of
several genes was impaired in cultured mast cells (CMCs) of
mi/mi mice due to a defective
transactivation ability of mutant MITF (mi-MITF). We
also found that mi/mi CMCs did not
express a receptor (MC1R) for 
-melanocyte-stimulating hormone. The
overexpression of the wild-type (+/+) MITF but not
mi-MITF normalized the expression of the MC1R in
mi/mi CMCs, indicating the involvement of
+-MITF in the MC1R gene expression. Next, we analyzed the promoter
region of the MC1R gene by the transient cotransfection assay. The
luciferase construct under the control of the MC1R promoter and the
cDNA-encoding +-MITF or mi-MITF were cotransfected into
NIH/3T3 fibroblasts. The cotransfection of +-MITF but not
mi-MITF increased the luciferase activity. There were
five CANNTG motifs recognized by bHLH-Zip-type transcription factors in
the cloned promoter region. We found +-MITF bound two of five CANNTG
motifs, and both motifs were essential for the transactivation of the
MC1R gene by +-MITF. These results indicated that +-MITF directly
transactivated the MC1R gene through these two
motifs.
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