Indirect recruitment of the signalling adaptor Shc to the fibroblast growth factor receptor 2 (FGFR2).
The adaptor protein Shc (Src homology and collagen-containing protein) plays an important role in the activation of signalling pathways downstream of RTKs (receptor tyrosine kinases) regulating diverse cellular functions, such as differentiation, adhesion, migration and mitogenesis. Despite being phosphorylated downstream of members of the FGFR (fibroblast growth factor receptor) family, a direct interaction of Shc with this receptor family has not been described to date. Various studies have suggested potential binding sites for the Shc PTB domain (phosphotyrosine-binding domain) and/or the SH2 (Src homology 2) domain on FGFR1, but no interaction of full-length Shc with these sites has been reported in vivo. In the present study, we investigated the importance of the SH2 domain and the PTB domain in recruitment of Shc to FGFR2(IIIc) to characterize the interaction of these two proteins. Confocal microscopy revealed extensive co-localization of Shc with FGFR2. The PTB domain was identified as the critical component of Shc which mediates membrane localization. Results from FLIM (fluorescence lifetime imaging microscopy) revealed that the interaction between Shc and FGFR2 is indirect, suggesting that the adaptor protein forms part of a signalling complex containing the receptor. We identified the non-RTK Src as a protein which potentially mediates the formation of such a ternary complex. Although an interaction between Src and Shc has been described previously, in the present study we implicate the Shc SH2 domain as a novel mediator of this association. The recruitment of Shc to FGFR2 via an indirect mechanism provides new insight into the regulation of protein assembly and activation of various signalling pathways downstream of this RTK.
Schüller
Annika C
AC
Department of Biochemistry and Molecular Biology, Darwin Building, Gower Street, London WC1E6BT, UK.
Ahmed
Zamal
Z
Levitt
James A
JA
Suen
Kin M
KM
Suhling
Klaus
K
Ladbury
John E
JE
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D017930:Genes, Reporter; D006801:Humans; D007425:Intracellular Membranes / Q000502:physiology; D007668:Kidney / Q000196:embryology; D016716:PC12 Cells; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D051381:Rats; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D051497:Receptor, Fibroblast Growth Factor, Type 2 / Q000235:genetics / Q000502:physiology*; D055765:Shc Signaling Adaptor Proteins / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection
D018076:DNA, Complementary; D055765:Shc Signaling Adaptor Proteins; C496353:FGFR2 protein, human; D020794:Receptor Protein-Tyrosine Kinases; D051497:Receptor, Fibroblast Growth Factor, Type 2