Abstract
Despite much interest in vascular endothelial growth factor (VEGF) and its receptors (VEGFRs -1 and -2), VEGF-induced signalling cascades remain incompletely defined. Attempts to assign individual responses to a particular receptor have used either transfected cell lines, receptor-specific growth factors or antisense oligonucleotides. Such studies have attributed the majority of VEGF-induced responses to activation of VEGFR-2. As a consequence of poor growth factor-induced VEGFR-1 autophosphorylation however, observations from these studies may instead reflect the relative activation of the two receptors. We have generated novel chimeric VEGF receptors in which the dimerization domain of the B subunit of DNA gyrase is fused to the cytoplasmic domain of VEGFRs -1 and -2. When expressed in porcine aortic endothelial cells, both chimeric VEGFR-1 and -2 autophosphorylate in response to addition of the small-molecule dimerizing agent, coumermycin. Once activated, both receptors induce downstream signalling cascades, exemplified here by the activation of MAPK, PLCĪ³ and PKB/Akt. Furthermore, we demonstrate that the Y1175 residue of VEGFR-2 is essential for the activation of PLCĪ³ mediated by this chimeric receptor. In contrast to previous reports which show a limited ability of VEGFR-1 to mediate signalling cascades, we show that once sufficiently activated, VEGFR-1 signals in a similar manner to VEGFR-2 in endothelial cells.
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Abbreviations
- BAEC:
-
bovine aortic endothelial cells
- ERK:
-
extracellular regulated kinase
- GyrB:
-
B subunit of DNA gyrase
- HUVEC:
-
human umbilical vein endothelial cells
- PAE cells:
-
porcine aortic endothelial cells
- PI3 kinase:
-
phosphatidyl inositol 3ā²-kinase
- PLCĪ³:
-
phospholipase CĪ³
- VEGF:
-
vascular endothelial growth factor
- VEGFR-1:
-
VEGF receptor 1
- VEGFR-2:
-
VEGF receptor 2
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Acknowledgements
We are grateful to Lena Claesson-Welsh, Rudbeck Laboratory, Uppsala, Sweden, for providing the VEGFR-1 cDNA and PAE cells. This work was supported in part by grants from the Wellcome Trust, British Heart Foundation, and Lister Institute for Preventive Medicine (SA Prigent).
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Knight, E., Warner, A., Maxwell, A. et al. Chimeric VEGFRs are activated by a small-molecule dimerizer and mediate downstream signalling cascades in endothelial cells. Oncogene 19, 5398ā5405 (2000). https://doi.org/10.1038/sj.onc.1203915
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DOI: https://doi.org/10.1038/sj.onc.1203915
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