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Originally published In Press as doi:10.1074/jbc.M101626200 on May 29, 2001

J. Biol. Chem., Vol. 276, Issue 30, 28147-28154, July 27, 2001
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Involvement of ITF2 in the Transcriptional Regulation of Melanogenic Genes*

Minao FurumuraDagger , S. Brian PotterfDagger , Kazutomo ToyofukuDagger , Jun MatsunagaDagger , Jacqueline Muller§, and Vincent J. HearingDagger

From the Dagger  Pigment Cell Biology Section, Laboratory of Cell Biology, NCI, National Institutes of Health, Bethesda, Maryland 20892 and the § Division of Virology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892

In response to agouti signal protein, melanocytes switch from producing eumelanin to pheomelanin concomitant with the down-regulation of melanogenic gene transcription. We previously reported that a ubiquitous basic helix-loop-helix transcription factor, known as ITF2, is up-regulated during this switch, and we now report that treatment of melanocytes with melanocyte-stimulating hormone down-regulates expression of ITF2. To more fully characterize the involvement of ITF2 in regulating melanogenic gene transcription, ITF2 sense or antisense constructs were introduced into melan-a melanocytes. Gene and protein expression analyses and luciferase reporter assays using promoters from melanogenic genes showed that up-regulation of ITF2 suppressed melanogenic gene expression as well as the expression of Mitf, a melanocyte-specific transcription factor. In addition, stable ITF2 sense transfectants had significant reductions in pigmentation and a less dendritic phenotype compared with mock transfectants. In contrast, ITF2 antisense-transfected melanocytes were more pigmented and more dendritic. These results demonstrate that up-regulation of ITF2 during the pheomelanin switch is functionally significant and reveal that differential expression of a ubiquitous basic helix-loop-helix transcription factor can modulate expression of melanogenic genes and the differentiation of melanocytes.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Laboratory of Cell Biology, Bldg. 37, Rm. 1B25, NCI, National Institutes of Health, Bethesda, MD 20892. Tel.: 301-496-1564; Fax: 301-402-8787; E-mail: hearingv@nih.gov.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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