Atypical receptor-mediated signal transduction events in the EGF-dependent growth-inhibited cell line, MDA-468.
It is now generally considered that early signalling from tyrosine kinases that induce mitogenesis is initiated through the formation of heteromeric complexes consisting of the autophosphorylated tyrosine kinase and a number of tyrosylphosphorylated proteins, including phospholipase C-gamma (PLC-gamma) and GTPase activating protein (GAP). However, since much of this work has been performed on proliferative, chimeric cell lines expressing heterologous receptor molecules, we examined the nature of the epidermal growth factor receptor (EGFR) signalling complex formation in the human breast cancer cell line, MDA-468. This cell line has an amplified, native EGFR gene, correspondingly overexpresses the EGFR, and its growth in culture is inversely related to the EGF concentration. Our results indicate that in MDA-468 cells, both the EGFR and PLC-gamma are phosphorylated on tyrosine residues and can be co-immunoprecipitated. This occurs at both high and low EGF concentrations regardless of the proliferative endpoint. The molecular association is correlated with a significant increase in total inositol phosphates formed in response to the growth factor treatment. In contrast, however, there is no evidence that GAP is either phosphorylated on tyrosine residues or forms a complex with the activated EGFR in EGF-treated MDA-468 cells. These observations suggest that as a model for growth factor action, the formation of heteromeric protein signalling complexes may demonstrate considerable diversity depending upon both cell type and physiology.
Church
J G
JG
Terry Fox Cancer Research Laboratories, Division of Basic Medical Sciences, Memorial University of Newfoundland, St. John's, Canada.
Richardson
V J
VJ
Lockwood
A G
AG
D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D004815:Epidermal Growth Factor / Q000494:pharmacology / Q000502:physiology*; D066246:ErbB Receptors / Q000502:physiology*; D020690:GTPase-Activating Proteins; D006801:Humans; D010716:Phosphatidylinositols / Q000378:metabolism; D010766:Phosphorylation; D011233:Precipitin Tests; D011506:Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000473:pathology; D010738:Type C Phospholipases / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D020690:GTPase-Activating Proteins; D010716:Phosphatidylinositols; D011506:Proteins; D014443:Tyrosine; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D010738:Type C Phospholipases