Sos1 rapidly associates with Grb2 and is hypophosphorylated when complexed with the EGF receptor after EGF stimulation.
The Son of sevenless (Sos) protein, a guanine nucleotide exchange factor for ras proteins, appears to play a central role in signalling between protein tyrosine kinase receptors and ras. The C-terminal region of Sos binds an adaptor protein, Grb2, which in turn binds to activated receptors including the EGF receptor (EGFR). Although the Sos protein is rapidly phosphorylated following cytokine stimulation, there is no evidence that this alters the enzymatic activity of Sos for ras proteins. Therefore, we investigated whether the ability of Sos1 to form complexes with Grb2 and with the EGF receptor (EGFR) changes following EGF stimulation, as a possible mechanism for regulating Sos activity. In contrast to earlier findings, we find that both the association and dissociation of Sos1 with Grb2 is responsive to EGF. Whilst the association of Sos1 and Grb2 following EGF stimulation is not cell type specific, we find that it is dependent on cell density and that the response to EGF differs to that induced by NGF. We find that following EGF stimulation, the Sos1 protein associated with the EGFR is markedly less phosphorylated than the majority of the Sos1 within the cell and there was reduced binding of Grb2 with phosphorylated Sos1 protein in a direct binding assay. A time course analysis showed that Sos1 dissociates from the EGFR more rapidly than does Grb2 following EGF stimulation. Collectively our findings are consistent with the notion that the phosphorylation of Sos1 affects its ability to complex with the EGFR and Grb2.
Hu
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Trescowthick Research Centre, Peter MacCallum Cancer Institute, Victoria, Australia.
Bowtell
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D016475:3T3 Cells; D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D066246:ErbB Receptors / Q000187:drug effects / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D051380:GRB2 Adaptor Protein; D006801:Humans; D051379:Mice; D009414:Nerve Growth Factors / Q000494:pharmacology; D016716:PC12 Cells; D010766:Phosphorylation; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D012097:Repressor Proteins / Q000378:metabolism*; D020837:SOS1 Protein; D015398:Signal Transduction
D048868:Adaptor Proteins, Signal Transducing; D005656:Fungal Proteins; D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; C495949:Grb2 protein, mouse; C495950:Grb2 protein, rat; D009414:Nerve Growth Factors; D011506:Proteins; D012097:Repressor Proteins; D020837:SOS1 Protein; D004815:Epidermal Growth Factor; D066246:ErbB Receptors