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Volume 272, Number 17, Issue of April 25, 1997 pp. 11026-11034
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Down-regulation of Epidermal Growth Factor Receptors by Nerve Growth Factor in PC12 Cells Is p140trk-, Ras-, and Src-dependent

(Received for publication, October 8, 1996, and in revised form, February 17, 1997)

Philip Lazarovici , Mari Oshima , Davidit Shavit , Makoto Shibutani , Hao Jiang , Mariam Monshipouri , Donald Fink § , Vilen Movsesyan and Gordon Guroff

From the Section on Growth Factors, NICHD, National Institutes of Health, Bethesda, Maryland 20892 and § Center of Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892

Nerve growth factor (NGF) treatment causes a profound down-regulation of epidermal growth factor receptors during the differentiation of PC12 cells. This process is characterized by a progressive decrease in epidermal growth factor (EGF) receptor level measured by 125I-EGF binding, tyrosine phosphorylation, and Western blotting. Treatment of the cells with NGF for 5 days produces a 95% reduction in the amount of [35S]methionine-labeled EGF receptors. This down-regulation does not occur in PC12nnr5 cells, which lack the p140trk NGF receptor. However, in PC12nnr5 cells stably transfected with p140trk, the NGF-induced heterologous down-regulation of EGF receptors is reconstituted in part. NGF-induced heterologous down-regulation, but not EGF-induced homologous down-regulation of EGF receptors, is blocked in Ras- and Src-dominant-negative PC12 cells. Treatment with either pituitary adenylate cyclase-activating peptide (PACAP) or staurosporine stimulates neurite outgrowth in PC12 cell variants, but neither induces down-regulation of EGF receptors. NGF treatment of PC12 cells in suspension induces down-regulation of EGF receptors in the absence of neurite outgrowth. These results strongly suggest a p140trk-, Ras- and Src-dependent mechanism of NGF-induced down-regulation of EGF receptors and separate this process from NGF-induced neurite outgrowth in PC12 cells.


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