NCK and PAK Participate in the Signaling Pathway by Which Vascular Endothelial Growth Factor Stimulates the Assembly of Focal Adhesions

Stoletov, Konstantin; Ratcliffe, Kirsty; Spring, Simone C.; Terman, Bruce I.

doi:10.1074/jbc.m009720200

Cited by 57 publications

(59 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recent reports have implicated Src and FAK in signal transduction linked to cell migration. [31][32][33][34] We thus prepared cell lysates from HUVEC stimulated with VEGF in the presence or absence of Sema-ECD. After SDS-PAGE, the resolved proteins were immunoblotted with a mAb to Src phosphorylated tyrosine 416 (pSrc-Y 416 ).…”

Section: Resultsmentioning

confidence: 99%

Recombinant semaphorin 6A-1 ectodomain inhibits in vivo growth factor and tumor cell line-induced angiogenesis

Mohanraj

Wu²,

Alvarez³

et al. 2005

Cancer Biology & Therapy

View full text Add to dashboard Cite

ACKNOWLEDGEMENTSWe thank CHTN and NDRI tissues and associated pathology reports. We thank Muralidhara Padigaru for genomic mining, Ramakrishnan Sundaram for performing the nude mouse angiogenesis assays; Dinesh Raturi, Pino Luan for protein production and technical assistance. We thank Dr. Stephen Strittmatter, Yale University School of Medicine for his support in providing reagents and also for critical reading of the manuscript. Research PaperRecombinant Semaphorin 6A-1 Ectodomain Inhibits In Vivo Growth Factor and Tumor Cell Line-Induced Angiogenesis ABSTRACTThe Semaphorins are a large family of transmembrane, GPI-anchored and secreted proteins that play an important role in neuronal and endothelial cell guidance. A human gene related to the class 6 Semaphorin family, Semaphorin 6A-1 (Sema 6A-1) was identified by homology-based genomic mining. Recent implication of Sema 3 family members in tumor angiogenesis and our expression analysis of Sema 6A-1 suggested that class 6 Semaphorin might effect tumor neovascularization. The mRNA expression of Sema 6A-1 was elevated in several renal tumor tissue samples relative to adjacent nontumor tissue samples from the same patient. Sema 6A-1 transcript was also expressed in the majority of renal clear cell carcinoma (RCC) cell lines and to a lesser extent in endothelial cells. To test the role of Sema 6A-1 in tumor angiogenesis, we engineered, expressed and purified the Sema 6A-1 soluble extracellular domain (Sema-ECD). The purified Sema-ECD was screened in a variety of endothelial cell-based assays both in vitro and in vivo. In vitro, Sema-ECD blocked VEGF-mediated endothelial cell migration. These effects were explained in part by our observation in endothelial cells that Sema-ECD inhibited VEGF-mediated Src, FAK and ERK phosphorylation. In vivo, mouse Matrigel assays demonstrated that the intraperitoneal administration of recombinant Sema-ECD inhibited both bFGF/VEGF and tumor cell line-induced neovascularization. These findings reveal a novel therapeutic utility for Sema 6A-1 (Sema-ECD) as an inhibitor of growth factor as well as tumor-induced angiogenesis.

show abstract

Section: Resultsmentioning

confidence: 99%

Recombinant semaphorin 6A-1 ectodomain inhibits in vivo growth factor and tumor cell line-induced angiogenesis

Mohanraj

Wu²,

Alvarez³

et al. 2005

Cancer Biology & Therapy

View full text Add to dashboard Cite

show abstract

“…Stoletov and colleagues have recently shown that VEGF stimulation recruited the Nck-Pak complex to focal adhesions, and the Nck-Pak complexed with FAK in human umbilical vein endothelial cells (HUVE). Introducing antisense oligonucleotides against Ncka into these cells inhibited VEGF-stimulated focal adhesion formation and migration (Stoletov et al, 2001). This ®nding suggests that Ncka has a speci®c role in VEGF signaling, since the 19-mer antisense oligonucleotide used by Stoletov et al (2001) unlikely cross reacted with Nckb mRNA and previous studies showed that both Ncka and Nckb are copresent in almost all cell types (Chen et al, , 2000, Nonetheless, it is important to test if down regulation of Nckb in these cells has any inhibitory e ects.…”

Section: Nck Links Cell Surface Receptors To the Actin Cytoskeletonmentioning

confidence: 99%

“…This suggests that Ncka and Nckb are functionally redundant during development. On the other hand, individual Nck can play speci®c roles in various cell types (Chen et al, , 2000Tu et al, 1998;Stoletov et al, 2001). Therefore, cell lines derived from various tissues of the Nck-knockout mice could further help our understanding of Nck function.…”

Section: Future Studiesmentioning

confidence: 99%

Nck/Dock: an adapter between cell surface receptors and the actin cytoskeleton

2001

View full text Add to dashboard Cite

In response to extracellular signals, cell surface receptors engage in connections with multiple intracellular signaling pathways, leading to the cellular responses such as survival, migration, proliferation and di erentiation. Thè pY?SH2/SH3?e ector' connection is a frequently used scheme by many cell surface receptors, in which SH2/SH3-containing adapters connect protein tyrosine phosphorylation to a variety of downstream e ector pathways. Following the initial landmark ®nding that Grb2 adapter links the receptors to the Ras pathway leading to DNA synthesis, recent studies have revealed that the biological function of the SH2/SH3 adapter Nck/Dock is to link cell surface receptors to the actin cytoskeleton. For example, in the evolutionarily-conserved signaling network, GEF-Rac-Nck-Pak, Nck`®xes up' the interaction of Pak with its upstream activator, Rac. The activated Pak then regulates the cytoskeletal dynamics. The fact that the majority of the more than 20 Nck-SH3-associated e ectors are regulators of the actin cytoskeleton suggests that Nck/Dock regulates, via binding to distinct e ectors, various cell type-speci®c motogenic responses. This review focuses on our current understanding of Nck/Dock function. Due to the number and complexity of the terminologies used in this review, à Glossary of Terms' is provided to help reduce confusions. Oncogene (2001) 20, 6403 ± 6417.Keywords: Src-homology domains; adapters; tyrosine kinases; signal transduction; cytoskeleton The SH2/SH3 adapters The src-homology 2 (SH2) and src-homlogy 3 (SH3) domains are protein ± protein interaction modules, found in a variety of structurally and functionally distinct signaling molecules, including protein tyrosine kinases/phosphatases, lipid kinase/phosphatase, guanine nucleotide exchange factors/GTPase activation proteins, docking proteins/adapters, cytoskeletal binding proteins and even transcription factors (Pawson, 1995;Mayer, 1999). Except for the terminology, SH2 and SH3 domains share no other common features regarding their structures, binding speci®city, and cellular targets. SH2 domains enable their host proteins to form complexes with phosphotyrosine (pY) proteins by binding to pY residues within these proteins. The stability and speci®city of the binding also depend upon the additional three to six amino acid residues immediately carboxyl to the pY residue, pY-(X) 3 ± 6 . Therefore, SH2 domains from di erent host molecules bind di erent pY-containing peptides. SH3 domains bind proline-rich segments of target molecules with a minimum consensus of P-x-x-P (Mayer, 2001). Each proline is often preceded by an aliphatic amino acid residue, and the aliphatic-proline pair binds to a hydrophobic pocket on SH3 domains. A major di erence between the SH2-pY binding and SH3-PxxP interactions is that the former is always transiently induced by extracellular signals, whereas the latter is often, but not always, constitutive. For instance, the SH2 domain of the adapter protein Grb2 only binds to tyrosine phosphorylated/activated EGFR, whe...

show abstract

“…Paks have previously been shown to act downstream of basic fibroblast growth factor and integrins (18) during angiogenesis and to bind the adaptor protein, NCK, an interaction required for angiogenesis (15) and the correct formation of focal adhesions (19,20). PAK2 has also been shown to induce retraction of endothelial cells (17).…”

Section: Discussionmentioning

confidence: 99%

“…Alterations in PAK function in vitro have been shown to affect endothelial contractility, retraction, and migration (14)(15)(16)(17). PAKs are activated by the growth factors basic fibroblast growth factor and VEGF and have been implicated in angiogenesis (15,18) and formation of focal adhesions (19,20); however, there is very limited information available about the functions of PAK proteins in vivo (21).…”

mentioning

confidence: 99%

pak2a mutations cause cerebral hemorrhage in redhead zebrafish

Buchner

Su²,

Yamaoka

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The zebrafish is a powerful model for studying vascular development, demonstrating remarkable conservation of this process with mammals. Here, we identify a zebrafish mutant, redhead (rhd mi149 ), that exhibits embryonic CNS hemorrhage with intact gross development of the vasculature and normal hemostatic function. We show that the rhd phenotype is caused by a hypomorphic mutation in p21-activated kinase 2a (pak2a). PAK2 is a kinase that acts downstream of the Rho-family GTPases CDC42 and RAC and has been implicated in angiogenesis, regulation of cytoskeletal structure, and endothelial cell migration and contractility among other functions. Correction of the Pak2a-deficient phenotype by Pak2a overexpression depends on kinase activity, implicating Pak2 signaling in the maintenance of vascular integrity. Rescue by an endothelial-specific transgene further suggests that the hemorrhage seen in Pak2a deficiency is the result of an autonomous endothelial cell defect. Reduced expression of another PAK2 ortholog, pak2b, in Pak2a-deficient embryos results in a more severe hemorrhagic phenotype, consistent with partially overlapping functions for these two orthologs. These data provide in vivo evidence for a critical function of Pak2 in vascular integrity and demonstrate a severe disease phenotype resulting from loss of Pak2 function.␤-pix ͉ CNS ͉ endothelial cell ͉ p21-activated kinase ͉ vasculature

show abstract

NCK and PAK Participate in the Signaling Pathway by Which Vascular Endothelial Growth Factor Stimulates the Assembly of Focal Adhesions

Cited by 57 publications

References 67 publications

Recombinant semaphorin 6A-1 ectodomain inhibits in vivo growth factor and tumor cell line-induced angiogenesis

Recombinant semaphorin 6A-1 ectodomain inhibits in vivo growth factor and tumor cell line-induced angiogenesis

Nck/Dock: an adapter between cell surface receptors and the actin cytoskeleton

pak2a mutations cause cerebral hemorrhage in redhead zebrafish

Contact Info

Product

Resources

About