Angiogenesis

, Volume 15, Issue 2, pp 187–197 | Cite as

c-ABL modulates MAP kinases activation downstream of VEGFR-2 signaling by direct phosphorylation of the adaptor proteins GRB2 and NCK1

  • Francesca Anselmi
  • Maurizio Orlandini
  • Marina Rocchigiani
  • Caterina De Clemente
  • Ahmad Salameh
  • Claudia Lentucci
  • Salvatore Oliviero
  • Federico Galvagni
Original Paper

Abstract

Vascular Endothelial Growth Factor-A (VEGF-A) is a key molecule in normal and tumor angiogenesis. This study addresses the role of c-ABL as a novel downstream target of VEGF-A in primary Human Umbilical Vein Endothelial Cells (HUVEC). On the basis of immunoprecipitation experiments, in vitro kinase assay and RNA interference, we demonstrate that VEGF-A induces the c-ABL kinase activity through the VEGF Receptor-2/Phosphatidylinositol-3-Kinase pathway. By treating HUVEC with the specific tyrosine kinase inhibitor STI571 and over-expressing a dominant negative c-ABL mutant, we show that the VEGF-A-activated c-ABL reduces the amplitude of Mitogen-Activated Protein Kinases (ERK1/2, JNKs and p38) activation in a dose-dependent manner by a negative feedback mechanism. By analysis of the adaptor proteins NCK1 and GRB2 mutants we further show that the negative loop on p38 is mediated by c-ABL phosphorylation at tyrosine 105 of the adaptor protein NCK1, while the phosphorylation at tyrosine 209 of GRB2 down-modulates ERK1/2 and JNKs signaling. These findings suggest that c-ABL function is to establish a correct and tightly controlled response of endothelial cells to VEGF-A during the angiogenic process.

Keywords

Angiogenesis VEGF Signal transduction NCK ABL 

Supplementary material

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Francesca Anselmi
    • 1
  • Maurizio Orlandini
    • 1
  • Marina Rocchigiani
    • 1
  • Caterina De Clemente
    • 1
  • Ahmad Salameh
    • 1
  • Claudia Lentucci
    • 1
  • Salvatore Oliviero
    • 1
  • Federico Galvagni
    • 1
  1. 1.Dipartimento di BiotecnologieUniversità di SienaSienaItaly

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