IQGAP1-dependent signaling pathway regulates endothelial cell proliferation and angiogenesis.
BACKGROUND
Vascular endothelial growth factor receptor-2 (VEGFR-2) signaling is an obligate requirement for normal development and pathological angiogenesis such as cancer and age-related macular degeneration. Although autophosphorylation of tyrosine 1173 (Y1173) of VEGFR-2 is considered a focal point for its angiogenic signal relay, however, the mechanism of phosphorylation of Y1173, signaling proteins that are recruited to this residue and their role in angiogenesis is not fully understood.


RESULTS
In this study we demonstrate that c-Src kinase directly through its Src homology 2 (SH2) domain and indirectly via c-Cbl binds to phospho-Y1057 of VEGFR-2. Activation of c-Src kinase by a positive feedback mechanism phosphorylates VEGFR-2 at multi-docking site, Y1173. c-Src also catalyzes tyrosine phosphorylation of IQGAP1 and acts as an adaptor to bridge IQGAP1 to VEGFR-2. In turn, IQGAP1 activates b-Raf and mediates proliferation of endothelial cells. Silencing expression of IQGAP1 and b-Raf revealed that their activity is essential for VEGF to stimulate angiogenesis in an in vivo angiogenesis model of chicken chorioallantoic membrane (CAM).


CONCLUSIONS
Angiogenesis contributes to the pathology of numerous human diseases ranging from cancer to age-related macular degeneration. Determining molecular mechanism of tyrosine phosphorylation of VEGFR-2 and identification of molecules that are relaying its angiogenic signaling may identify novel targets for therapeutic intervention against angiogenesis-associated diseases. Our study shows that recruitment and activation of c-Src by VEGFR-2 plays a pivotal role in relaying angiogenic signaling of VEGFR-2; it phosphorylates VEGFR-2 at Y1173, facilitates association and activation of IQGAP1 and other signaling proteins to VEGFR-2. IQGAP1-dependent signaling, in part, is critically required for endothelial cell proliferation, a key step in angiogenesis. Thus, Y1057 of VEGFR-2 serves to regulate VEGFR-2 function in a combinatorial manner by supporting both diversity of recruitment of angiogenic signaling proteins to VEGFR-2, and its ability to promote angiogenesis.
Meyer
Rosana D
RD
Department of Pathology and Ophthalmology, School of Medicine, Boston University, Boston, MA, USA.
Sacks
David B
DB
Rahimi
Nader
N
D000818:Animals; D015153:Blotting, Western; D049109:Cell Proliferation; D002645:Chickens; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D010766:Phosphorylation; D050721:Proto-Oncogene Proteins c-cbl / Q000378:metabolism*; D015398:Signal Transduction; D014443:Tyrosine / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism*; D020703:ras GTPase-Activating Proteins / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism*
C101203:IQ motif containing GTPase activating protein 1; D020703:ras GTPase-Activating Proteins; D014443:Tyrosine; D050721:Proto-Oncogene Proteins c-cbl; D040301:Vascular Endothelial Growth Factor Receptor-2; D019061:src-Family Kinases