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A more recent version of this article appeared on February 9, 2001
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M008776200v1
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Papers In Press, published online ahead of print November 8, 2000
J. Biol. Chem, 10.1074/jbc.M008776200
Submitted on September 26, 2000
Revised on November 8, 2000
Accepted on November 8, 2000

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 Liu, Li-Jie Gong, Li-Ying Guo, Yasuhiro Katagiri, Hao Jiang, Zhao-Yi Wang, Alfred C. Johnson, and Gordon Guroff

Laboratory of Molecular Biology, National Cancer Institute, NIH, Bethesda, MD 20892-4255

Corresponding Author: aj2e{at}nih.gov

Recently, we characterized the rat epidermal growth factor receptor (EGFR) promoter and demonstrated that a TCC repeat sequences is required for the down-regulation of EGFR by nerve growth factor (NGF) in PC12 cells. In this paper, 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 co-transfection assays. Gel mobility shift assays demonstrate that WT1 binds to the TCC repeat sequences of the rat EGFR promoter. Overexpression of WT1 resulted in the 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 and 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 found that antisense WT1 RNA could substantially reduce EGFR repression either in 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 WT1 may play an important role in the differentiation of nerve cells.


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