VEGF autoregulates its proliferative and migratory ERK1/2 and p38 cascades by enhancing the expression of DUSP1 and DUSP5 phosphatases in endothelial cells

Abstract

Fotsis T. VEGF autoregulates its proliferative and migratory ERK1/2 and p38 cascades by enhancing the expression of DUSP1 and DUSP5 phosphatases in endothelial cells. Am J Physiol Cell Physiol 297: C1477-C1489, 2009. First published September 9, 2009 doi:10.1152/ajpcell.00058.2009.-Vascular endothelial growth factor (VEGF) is a key angiogenic factor that regulates proliferation and migration of endothelial cells via phosphorylation of extracellular signal-regulated kinase-1/2 (ERK1/2) and p38, respectively. Here, we demonstrate that VEGF strongly induces the transcription of two dual-specificity phosphatase (DUSP) genes DUSP1 and DUSP5 in endothelial cells. Using fluorescence microscopy, fluorescence lifetime imaging (FLIM), and fluorescence cross-correlation spectroscopy (FCCS), we found that DUSP1/mitogen-activated protein kinases phosphatase-1 (MKP-1) was localized in both the nucleus and cytoplasm of endothelial cells, where it existed in complex with p38 (effective dissociation constant, K D eff , values of 294 and 197 nM, respectively), whereas DUSP5 was localized in the nucleus of endothelial cells in complex with ERK1/2 (K D eff 345 nM). VEGF administration affected differentially the K D eff values of the DUSP1/p38 and DUSP5/ERK1/2 complexes. Gain-of-function and lack-of-function approaches revealed that DUSP1/MKP-1 dephosphorylates primarily VEGF-phosphorylated p38, thereby inhibiting endothelial cell migration, whereas DUSP5 dephosphorylates VEGF-phosphorylated ERK1/2 inhibiting proliferation of endothelial cells. Moreover, DUSP5 exhibited considerable nuclear anchoring activity on ERK1/2 in the nucleus, thereby diminishing ERK1/2 export to the cytoplasm decreasing its further availability for activation. vascular endothelial growth factors; extracellular signal regulated kinase-1/2; p38; dual-specificity phosphatase 1/mitogen-activated protein kinase phosphatase-1; dual-specificity phosphatase-5 VASCULAR ENDOTHELIAL GROWTH FACTORS (VEGFs) are the most important regulators of vessel morphogenesis. Not only do they participate in the regulation of both vasculogenesis and angiogenesis, but they also are among the most important molecules involved in the pathogenesis of angiogenic diseases such as diabetic retinopathy and cancer (6). VEGFs are secreted dimeric glycoproteins of ϳ40 kDa, and in mammals the VEGF family consists of five members: VEFG-A, B, C, D and placental growth factor (PLGF) (6). Moreover, alternative splicing of several of the VEGF family members gives rise to isoforms with different biological activities. The VEGF ligands bind in an overlapping pattern to three receptor tyrosine kinases known as VEGF receptor-1, -2, and -3 (VEGFR1-3), as well as to coreceptors such as heparan sulfate proteoglycans and neuropilins (22). VEGFR2 promotes migration, proliferation, and differentiation of endothelial cells (ECs) being critical for the regulation of angiogenic sprouting (1). Indeed, during sprouting, tip cells need to acquire an invasive and motile phenotype, whereas ECs in the stalk exhibi...

Estrogen Defines the Dorsal-Ventral Limit of VEGF Regulation to Specify the Location of the Hemogenic Endothelial Niche

“…To confirm nascent E2 could nevertheless regulate VEGF activation, we examined embryos expressing GFP under the regulation of 1.2kb of the VEGF promoter (He and Chen, 2005) (Figure 6B); E2 strongly reduced VEGF activation, while ZK broadened the zone of expression. To determine if VEGF activity was also impacted, known transcriptional targets dll4 , dusp5 and notch1b (Bellou et al, 2009; Liu et al, 2003) (Figure 6C,D and Figure S7C) were assessed: inhibition of E2-signaling by ZK strongly enhanced expression of each VEGF target compared to controls, suggesting VEGF-mediated transcriptional regulation is actively antagonized by endogenous E2 in hemato-vascular development. Consistent with that finding, ZK treated embryos exhibited phenotypes reminiscent of those observed with VEGF overexpression in the ventral aspect of the AGM, including widening of the major vessels and ectopic or disorganized ISV sprouting (Figure S7A–B) (Lawson et al, 2002; Wiley et al, 2011).…”

Dynamic regulation of VEGF-inducible genes by an ERK-ERG-p300 transcriptional network

“…DUSP5, NRARP) regulate angiogenesis ( Fig. 9H) (Al-Greene et al, 2013;Bellou et al, 2009;Herbert et al, 2012;Iaccarino et al, 2005;Mirza et al, 2013;Moehler et al, 2008;Phng et al, 2009;Pin et al, 2012;Prahst et al, 2014;Toffoli et al, 2009;Zhou et al, 2011).…”

“…Several VEGF/MAPK/ERK-responsive genes have been characterized, including the immediate early gene early growth response 3 (EGR3) and the ERK phosphatase dual specificity phosphatase 5 (DUSP5) (Bellou et al, 2009;Kucharska et al, 2009). The transcriptional induction of EGR3 and DUSP5 (as measured by qRT-PCR analysis of unspliced pre-mRNA) largely mirrored that of MAPK/ERK activation and DLL4 transcription (Fig.…”

NADPH Oxidase Inhibition in Heart Failure Improved Vascular Function Associated with Changes in the Novel Genes Expression Revealed by Transcriptome Analysis

“…DUSP5 is a serine-threonine phosphatase and dephosphorylates vascular endothelial growth factor-phosphorylated ERK1/2 inhibiting proliferation of endothelial cells [17]. Intrinsic vascular smooth muscle cell signaling in the cascade of events leading to augmented total peripheral resistance in HF and the active myogenic response regulates the local increase in microvascular resistance.…”