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Originally published In Press as doi:10.1074/jbc.M008776200 on November 8, 2000

J. Biol. Chem., Vol. 276, Issue 7, 5068-5073, February 16, 2001
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The Wilms' Tumor Gene Product WT1 Mediates the Down-regulation of the Rat Epidermal Growth Factor Receptor by Nerve Growth Factor in PC12 Cells*

Xu-Wen LiuDagger , Li-Jie Gong§, Li-Ying GuoDagger , Yasuhiro KatagiriDagger ||, Hao JiangDagger **, Zhao-Yi WangDagger Dagger , Alfred C. Johnson§§¶¶, and Gordon Guroffdagger Dagger

From the Dagger  Section on Growth Factors and § Laboratory of Developmental and Molecular Immunity, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, Dagger Dagger  Division of Growth Regulation, Department of Medicine, Beth Israel-Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, and §§ Laboratory of Molecular Biology, Division of Basic Sciences, NCI, National Institutes of Health, Bethesda, Maryland 20892

Recently, we characterized the rat epidermal growth factor receptor (EGFR) promoter and demonstrated that TCC repeat sequences are required for the down-regulation of EGFR by nerve growth factor (NGF) in PC12 cells. In this study, we report that the Wilms' tumor gene product WT1, a zinc finger transcription factor, is able to enhance the activity of the rat EGFR promoter in cotransfection assays. Gel mobility shift assays demonstrate that WT1 binds to the TCC repeat sequences of the rat EGFR promoter. Overexpression of WT1 resulted in up-regulation of the expression levels of endogenous EGFR in PC12 cells. Interestingly, NGF down-regulated the expression levels of WT1 and EGFR in PC12 cells, but not in the p140trk-deficient variant PC12nnr5 cells or in cells expressing either dominant-negative Ras or dominant-negative Src. Most importantly, we evaluated the inhibitory effect of antisense WT1 RNA on EGFR expression, and we found that antisense WT1 RNA could substantially reduce EGFR repression in either histochemical staining study or immunoblot analysis. These results indicate that NGF-induced down-regulation of the EGFR in PC12 cells is mediated through WT1 and that WT1 may play an important role in the differentiation of nerve cells.

* 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.

dagger This article is dedicated to the memory of Dr. Gordon Guroff, Chief of Section on Growth Factors and Deputy Scientific Director of the National Institute of Child Health and Human Development, who died on July 9, 1999. During his long career, he enriched the field of neuroscience and all of us who had the privilege to know and work with him.

Present address: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.

|| Present address: American Red Cross, Holland Laboratory, Biochemistry Department, 15601 Crabbs Branch Way, Rockville, MD 20855.

** Present address: William T. Gossett Neurology Laboratories, Henry Ford Health Sciences Center, 1 Ford Place, 4D Research, Detroit, MI 48202.

¶¶ To whom correspondence should be addressed: Bldg. 37, Rm. 2D18, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892. Tel.: 301-496-3224; Fax: 301-402-1344; E-mail: aj2e@nih.gov.

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