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Volume 272, Number 1, Issue of January 3, 1997 pp. 503-509
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Functional Analysis of Microphthalmia-associated Transcription Factor in Pigment Cell-specific Transcription of the Human Tyrosinase Family Genes

(Received for publication, July 9, 1996, and in revised form, August 30, 1996)

Ken-ichi Yasumoto Dagger , Kouji Yokoyama Dagger , Kazuhiro Takahashi Dagger , Yasushi Tomita § and Shigeki Shibahara Dagger

From the Dagger  Department of Applied Physiology and Molecular Biology, Tohoku University School of Medicine, Aoba-ku, Sendai, Miyagi 980-77 and the § Department of Dermatology, Akita University School of Medicine, Akita 010, Japan

Tyrosinase, tyrosinase-related protein-1 (TRP-1), and TRP-2 are the enzymes involved in melanin biosynthesis and are preferentially expressed in pigment cells. Their human gene promoters share the 11-base pair M box containing a CATGTG motif, which was shown here to be bound in vitro by microphthalmia-associated transcription factor (MITF). Transient cotransfection analysis showed that MITF overexpression increased the expression of a reporter gene under the control of the human tyrosinase or TRP-1 gene promoter but not the TRP-2 promoter. The promoter activation caused by MITF is dependent on each CATGTG motif of the distal enhancer element, the M box, and the initiator E box of the tyrosinase gene and the TRP-1 M box. Furthermore, a truncated MITF lacking the carboxyl-terminal 125 amino acid residues transactivated the tyrosinase promoter less efficiently than did MITF, suggesting that MITF's carboxyl terminus contains a transcriptional activation domain, but unexpectedly such a truncated MITF remarkably transactivated the TRP-2 gene promoter. These results suggest that MITF is sufficient to direct pigment cell-specific transcription of the tyrosinase and TRP-1 genes but not the TRP-2 gene.


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