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  • Original Paper
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Isoforms of c-KIT differ in activation of signalling pathways and transformation of NIH3T3 fibroblasts

Oncogene volume 18, pages 5573–5581 (1999)Cite this article

Abstract

Alternate splicing of mRNA encoding c-KIT results in isoforms which differ in the presence or absence of four amino acids (GNNK) in the juxtamembrane region of the extracellular domain of the receptor. In this study we show that these isoforms of human c-KIT, expressed at similar levels in NIH3T3 cells, display differential effects on various attributes of transformation. The GNNK− isoform strongly promoted anchorage independent growth (colony formation in semi-solid medium), loss of contact inhibition (focus formation), and led to tumorigenicity in nude mice. In contrast, the GNNK+ isoform elicited colony formation but relatively poor focus formation and no tumorigenicity. Saturation binding analysis indicated that the isoforms do not differ significantly in their affinity for the KIT ligand, Steel Factor (SLF). Negligible ligand-independent receptor phosphorylation was observed in either case but, after ligand stimulation, the GNNK− isoform displayed more rapid and extensive tyrosine autophosphorylation and faster internalization. Both isoforms recruited the p85 subunit of phosphatidylinositol 3-kinase and led to similar phosphorylation of its downstream effector c-Akt, but the GNNK− isoform gave rise to more MAP kinase phosphorylation. Thus the c-KIT isoforms display different signalling characteristics and have different transforming activity in NIH3T3 cells.

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Acknowledgements

We thank Ly Nguyen for skilled technical assistance, Paul Sincock for help with confocal microscopy, Sonia Young for contributing binding data on PC12 cells, and Regan Forrest for carrying out preliminary signalling experiments. This work was supported by a grant from the National Health and Medical Research Council of Australia of which LK Ashman is a Senior Research Fellow, and by an Australian Postgraduate Award to G Caruana.

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Author notes

  1. Georgina Caruana

    Present address: Program in Molecular Biology and Cancer, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, M5G 1X5, Ontario, Canada

  2. Georgina Caruana: G Caruana and AC Cambareri contributed equally to this work

Authors and Affiliations

  1. Division of Haematology, Hanson Centre for Cancer Research, Institute of Medical and Veterinary Science, Box 14 Rundle Mall PO, Adelaide, SA 5000, Australia

    Georgina Caruana, Antony C Cambareri & Leonie K Ashman

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Caruana, G., Cambareri, A. & Ashman, L. Isoforms of c-KIT differ in activation of signalling pathways and transformation of NIH3T3 fibroblasts. Oncogene 18, 5573–5581 (1999). https://doi.org/10.1038/sj.onc.1202939

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