Expression of the v-crk oncogene product in PC12 cells results in rapid differentiation by both nerve growth factor- and epidermal growth factor-dependent pathways

Barbara L. Hempstead, Raymond B. Birge, Jorge E. Fajardo, Robert Glassman, Debbie Mahadeo, Rosemary Kraemer, Hidesaburo Hanafusa

Research output: Contribution to journalArticle

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Abstract

The transforming gene of the avian sarcoma virus CT10 encodes a fusion protein (p47gag-crk or v-Crk) containing viral Gag sequences fused to cellular sequences consisting primarily of Src homology regions 2 and 3 (SH2 and SH3 sequences). Here we report a novel function of v-Crk in the mammalian pheochromocytoma cell line, PC12, whereby stable expression of v-Crk induces accelerated differentiation, as assessed by induction of neurites following nerve growth factor (NGF) or basic fibroblast growth factor (bFGF) treatment compared with the effect in native PC12 cells. Surprisingly, however, these cells also develop extensive neurite processes after epidermal growth factor (EGF) stimulation, an event which is not observed in native PC12 cells. Following EGF or NGF stimulation of the v-CrkPC12 cells, the v-Crk protein itself became tyrosine phosphorylated within 1 min. Moreover, in A431 cells or TrkA-PC12 cells, which overexpress EGF receptors and TrkA, respectively, a GST-CrkSH2 fusion protein was indeed capable of binding these receptors in a phosphotyrosine-dependent manner, suggesting that v-Crk can directly couple to receptor tyrosine kinase pathways in PC12 cells. In transformed fibroblasts, v-Crk binds to specific tyrosine-phosphorylated proteins of p130 and paxillin. Both of these proteins are also complexed to v-Crk in PC12 cells, as evidenced by their coprecipitation with v-Crk in detergent lysates, suggesting that common effector pathways may occur in both cell types. However, whereas PC12 cellular differentiation can occur solely by overexpression of the v-Src or oncogenic Ras proteins, that induced by v-Crk requires a growth factor stimulatory signal, possibly in a two-step process.

Original languageEnglish (US)
Pages (from-to)1964-1971
Number of pages8
JournalMolecular and Cellular Biology
Volume14
Issue number3
StatePublished - Mar 1994
Externally publishedYes

Fingerprint

Oncogene Proteins
PC12 Cells
Nerve Growth Factor
Epidermal Growth Factor
Neurites
Oncogene Protein v-crk
Tyrosine
Proteins
Avian Sarcoma Viruses
Paxillin
ras Proteins
Phosphotyrosine
Receptor Protein-Tyrosine Kinases
Fibroblast Growth Factor 2
Oncogenes
Detergents
Intercellular Signaling Peptides and Proteins
Fibroblasts

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Expression of the v-crk oncogene product in PC12 cells results in rapid differentiation by both nerve growth factor- and epidermal growth factor-dependent pathways. / Hempstead, Barbara L.; Birge, Raymond B.; Fajardo, Jorge E.; Glassman, Robert; Mahadeo, Debbie; Kraemer, Rosemary; Hanafusa, Hidesaburo.

In: Molecular and Cellular Biology, Vol. 14, No. 3, 03.1994, p. 1964-1971.

Research output: Contribution to journalArticle

Hempstead, Barbara L. ; Birge, Raymond B. ; Fajardo, Jorge E. ; Glassman, Robert ; Mahadeo, Debbie ; Kraemer, Rosemary ; Hanafusa, Hidesaburo. / Expression of the v-crk oncogene product in PC12 cells results in rapid differentiation by both nerve growth factor- and epidermal growth factor-dependent pathways. In: Molecular and Cellular Biology. 1994 ; Vol. 14, No. 3. pp. 1964-1971.
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