Hepatocyte Growth Factor-induced Scatter of Madin-Darby Canine Kidney Cells Requires Phosphatidylinositol 3-Kinase

doi:10.1074/jbc.270.46.27780

Hepatocyte Growth Factor-induced Scatter of Madin-Darby Canine Kidney Cells Requires Phosphatidylinositol 3-Kinase (*)

From the (1) Molecular Oncology Laboratory, Royal Victoria Hospital, the Departments of Medicine,
(2) Oncology, and
(3) Biochemistry, McGill University, Montreal, Quebec H3A 1A1, Canada

¶ Senior scholar of the National Cancer Institute of Canada. To whom correspondence should be addressed:
Molecular Oncology Group, H5.21, 687 Pine Ave. West, Montreal, Quebec H3A 1A1, Canada.
Tel.: 514-842-1231, ext. 5834; Fax: 514-843-1478.

Abstract

Hepatocyte growth factor/scatter factor (HGF/SF) is a multifunctional cytokine that induces mitogenesis, motility, invasion, and morphogenesis of several epithelial and endothelial cell lines in culture. The receptor for HGF/SF has been identified as the Met tyrosine kinase. To investigate the signaling pathways that are involved in these events, we have generated chimeric receptors containing the extracellular domain of the colony-stimulating factor-1 (CSF-1) receptor fused to the transmembrane and intracellular domains of the Met receptor (MET). Madin-Darby canine kidney (MDCK) epithelial cells expressing the CSF-MET chimera dissociate and scatter in response to CSF-1. However, cells expressing a mutant CSF-MET receptor containing a phenylalanine substitution for tyrosine 1356 were unable to scatter or form branching tubules following stimulation with CSF-1. Tyrosine 1356 is essential for the recruitment of multiple substrates including the p85 subunit of PI3-kinase, phospholipase C, and Grb2. In this study, we have investigated the role of PI3-kinase and a downstream target of PI3-kinase, pp70, in the induction of MDCK cell scatter in response to HGF/SF. Our results demonstrate that following stimulation with HGF/SF, activation of PI3-kinase but not pp70 is essential for MDCK cell scatter.

Footnotes

↵§ Recipient of a fellowship from the Fonds de la Recherche en Santé du Québec.
↵* This research was supported by operating grants from the National Cancer Institute of Canada and the Medical Research Council of Canada (to M. P.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¹ The abbreviations used are:

HGF/SF

hepatocyte growth factor/scatter factor

CSF

colony-stimulating factor

MDCK

Madin-Darby canine kidney

PI3-kinase

phosphatidylinositol 3-kinase

ETOH

ethanol

PBS

phosphate-buffered saline

PIPES

1,4-piperazinediethanesulfonic acid

PDGF

platelet-derived growth factor

SH2

Src homology 2

PTB

phosphotyrosine binding.
↵² X. M. Yang and M. Park(1995) Lab. Invest., in press.
↵³ T. Fournier and M. Park, manuscript in preparation.
- Received June 15, 1995.
- Revision received September 8, 1995.