Abstract
Intercellular junctions mediate adhesion and communication between adjoining cells. Although formed by different molecules, tight junctions (TJs) and adherens junctions (AJs) are functionally and structurally linked, but the signalling pathways behind this interaction are unknown. Here we describe a cell-specific mechanism of crosstalk between these two types of structure. We show that endothelial VE-cadherin at AJs upregulates the gene encoding the TJ adhesive protein claudin-5. This effect requires the release of the inhibitory activity of forkhead box factor FoxO1 and the Tcf-4–β-catenin transcriptional repressor complex. Vascular endothelial (VE)-cadherin acts by inducing the phosphorylation of FoxO1 through Akt activation and by limiting the translocation of β-catenin to the nucleus. These results offer a molecular basis for the link between AJs and TJs and explain why VE-cadherin inhibition may cause a marked increase in permeability.
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References
Dejana, E. Endothelial cell–cell junctions: happy together. Nature Rev. Mol. Cell Biol. 5, 261–270 (2004).
Gumbiner, B. M. Regulation of cadherin-mediated adhesion in morphogenesis. Nature Rev. Mol. Cell Biol. 6, 622–634 (2005).
Furuse, M. & Tsukita, S. Claudins in occluding junctions of humans and flies. Trends Cell Biol. 16, 181–188 (2006).
Nitta, T. et al. Size-selective loosening of the blood–brain barrier in claudin-5-deficient mice. J. Cell Biol. 161, 653–660 (2003).
Ikenouchi, J., Umeda, K., Tsukita, S., Furuse, M. & Tsukita, S. Requirement of ZO-1 for the formation of belt-like adherens junctions during epithelial cell polarization. J. Cell Biol. 176, 779–786 (2007).
Miyoshi, J. & Takai, Y. Molecular perspective on tight-junction assembly and epithelial polarity. Adv. Drug Deliv. Rev. 57, 815–855 (2005).
Ohsugi, M., Larue, L., Schwarz, H. & Kemler, R. Cell-junctional and cytoskeletal organization in mouse blastocysts lacking E-cadherin. Dev. Biol. 185, 261–271 (1997).
Behrens, J., Birchmeier, W., Goodman, S. L. & Imhof, B. A. Dissociation of Madin–Darby canine kidney epithelial cells by the monoclonal antibody anti-arc-1: mechanistic aspects and identification of the antigen as a component related to uvomorulin. J. Cell Biol. 101, 1307–1315 (1985).
Carmeliet, P. et al. Targeted deficiency or cytosolic truncation of the VE-cadherin gene in mice impairs VEGF-mediated endothelial survival and angiogenesis. Cell 98, 147–157 (1999).
Corada, M. et al. Vascular endothelial-cadherin is an important determinant of microvascular integrity in vivo. Proc. Natl Acad. Sci. USA 96, 9815–9820 (1999).
Liebner, S., Cavallaro, U. & Dejana, E. The multiple languages of endothelial cell-to-cell communication. Arterioscler. Thromb. Vasc. Biol. 26, 1431–1438 (2006).
Wheelock, M. J. & Johnson, K. R. Cadherin-mediated cellular signaling. Curr. Opin. Cell Biol. 15, 509–514 (2003).
Lampugnani, M. G. et al. Contact inhibition of VEGF-induced proliferation requires vascular endothelial cadherin, β-catenin, and the phosphatase DEP-1/CD148. J. Cell Biol. 161, 793–804 (2003).
Xiao, K. et al. Mechanisms of VE-cadherin processing and degradation in microvascular endothelial cells. J. Biol. Chem. 278, 19199–19208 (2003).
Lampugnani, M. G. et al. VE-cadherin regulates endothelial actin activating Rac and increasing membrane association of Tiam. Mol. Biol. Cell 13, 1175–1189 (2002).
Crosby, C. V. et al. VE-cadherin is not required for the formation of nascent blood vessels but acts to prevent their disassembly. Blood 105, 2771–2776 (2005).
Gavard, J. & Gutkind, J. S. VEGF controls endothelial-cell permeability by promoting the β-arrestin-dependent endocytosis of VE-cadherin. Nature Cell Biol. 8, 1223–1234 (2006).
Daly, C. et al. Angiopoietin-1 modulates endothelial cell function and gene expression via the transcription factor FKHR (FOXO1). Genes Dev. 18, 1060–1071 (2004).
Burgering, B. M. & Kops, G. J. Cell cycle and death control: long live Forkheads. Trends Biochem. Sci. 27, 352–360 (2002).
Potente, M. et al. Involvement of Foxo transcription factors in angiogenesis and postnatal neovascularization. J. Clin. Invest 115, 2382–2392 (2005).
Brunet, A. et al. Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 96, 857–868 (1999).
Zhang, X. et al. Phosphorylation of serine 256 suppresses transactivation by FKHR (FOXO1) by multiple mechanisms. Direct and indirect effects on nuclear/cytoplasmic shuttling and DNA binding. J. Biol. Chem. 277, 45276–45284 (2002).
Gilley, J., Coffer, P. J. & Ham, J. FOXO transcription factors directly activate bim gene expression and promote apoptosis in sympathetic neurons. J. Cell Biol. 162, 613–622 (2003).
Paik, J. H. et al. FoxOs are lineage-restricted redundant tumor suppressors and regulate endothelial cell homeostasis. Cell 128, 309–323 (2007).
Lee, S. et al. Autocrine VEGF signaling is required for vascular homeostasis. Cell 130, 691–703 (2007).
Sini, P. et al. Inhibition of multiple vascular endothelial growth factor receptors (VEGFR) blocks lymph node metastases but inhibition of VEGFR-2 is sufficient to sensitize tumor cells to platinum-based chemotherapeutics. Cancer Res. 68, 1581–1592 (2008).
Faivre, S., Demetri, G., Sargent, W. & Raymond, E. Molecular basis for sunitinib efficacy and future clinical development. Nature Rev. Drug Discov. 6, 734–745 (2007).
Chen, J. et al. Akt1 regulates pathological angiogenesis, vascular maturation and permeability in vivo. Nature Med. 11, 1188–1196 (2005).
Essers, M. A. et al. Functional interaction between β-catenin and FOXO in oxidative stress signaling. Science 308, 1181–1184 (2005).
Clevers, H. Wnt/β-catenin signaling in development and disease. Cell 127, 469–480 (2006).
Vleminckx, K., Kemler, R. & Hecht, A. The C-terminal transactivation domain of β-catenin is necessary and sufficient for signaling by the LEF-1/β-catenin complex in Xenopus laevis. Mech. Dev. 81, 65–74 (1999).
Quasnichka, H. et al. Regulation of smooth muscle cell proliferation by β-catenin/T-cell factor signaling involves modulation of cyclin D1 and p21 expression. Circ. Res. 99, 1329–1337 (2006).
Van Itallie, C. M. & Anderson, J. M. Claudins and epithelial paracellular transport. Annu. Rev. Physiol 68, 403–429 (2006).
Dimmeler, S. & Zeiher, A. M. Akt takes center stage in angiogenesis signaling. Circ. Res. 86, 4–5 (2000).
Seoane, J., Le, H. V., Shen, L., Anderson, S. A. & Massagué, J. Integration of Smad and forkhead pathways in the control of neuroepithelial and glioblastoma cell proliferation. Cell 117, 211–223 (2004).
Asada, S. et al. Mitogen-activated protein kinases, Erk and p38, phosphorylate and regulate Foxo1. Cell Signal. 19, 519–527 (2007).
Dejana, E., Taddei, A. & Randi, A. M. Foxs and Ets in the transcriptional regulation of endothelial cell differentiation and angiogenesis. Biochim. Biophys. Acta 1775, 298–312 (2007).
Lampugnani, M. G. et al. Cell confluence regulates tyrosine phosphorylation of adherens junction components in endothelial cells. J. Cell Sci. 110, 2065–2077 (1997).
Roura, S., Miravet, S., Piedra, J., Garcia de, H. A. & Dunach, M. Regulation of E-cadherin/Catenin association by tyrosine phosphorylation. J. Biol. Chem. 274, 36734–36740 (1999).
Huber, A. H. & Weis, W. I. The structure of the β-catenin/E-cadherin complex and the molecular basis of diverse ligand recognition by β-catenin. Cell 105, 391–402 (2001).
Zanetta, L. et al. Downregulation of vascular endothelial-cadherin expression is associated with an increase in vascular tumor growth and hemorrhagic complications. Thromb. Haemost. 93, 1041–1046 (2005).
Cattelino, A. et al. The conditional inactivation of the β-catenin gene in endothelial cells causes a defective vascular pattern and increased vascular fragility. J. Cell Biol. 162, 1111–1122 (2003).
Dull, T. et al. A third-generation lentivirus vector with a conditional packaging system. J. Virol. 72, 8463–8471 (1998).
Pear, W. S., Nolan, G. P., Scott, M. L. & Baltimore, D. Production of high-titer helper-free retroviruses by transient transfection. Proc. Natl Acad. Sci. USA 90, 8392–8396 (1993).
Spagnuolo, R. et al. Gas1 is induced by VE-cadherin and vascular endothelial growth factor and inhibits endothelial cell apoptosis. Blood 103, 3005–3012 (2004).
Grueneberg, D. A. et al. A functional screen in human cells identifies UBF2 as an RNA polymerase II transcription factor that enhances the β-catenin signaling pathway. Mol. Cell Biol. 23, 3936–3950 (2003).
Furuyama, T., Nakazawa, T., Nakano, I. & Mori, N. Identification of the differential distribution patterns of mRNAs and consensus binding sequences for mouse DAF-16 homologues. Biochem. J. 349, 629–634 (2000).
Vecchi, M. et al. Gene expression analysis of early and advanced gastric cancers. Oncogene 26, 4284–4294 (2007).
Cartharius, K. et al. MatInspector and beyond: promoter analysis based on transcription factor binding sites. Bioinformatics 21, 2933–2942 (2005).
Nakae, J. et al. The forkhead transcription factor Foxo1 regulates adipocyte differentiation. Dev. Cell 4, 119–129 (2003).
Acknowledgements
We thank Barbara Felice for help in the transcription-factor-binding-site analysis. This work was supported by the Associazione Italiana per la Ricerca sul Cancro, Association for International Cancer Research, the European Community (Integrated Project Contract No LSHG-CT-2004-503573; NoE MAIN 502935; NoE EVGN 503254; EUSTROKE and OPTISTEM Networks), Istituto Superiore di Sanita', Italian Ministry of Health, MIUR (COFIN prot: 2006058482_002), Fondation Leducq Transatlantic Network of Excellence. C.G. is supported by an AIRC-SISAL fellowship.
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E.D. and A.T. planned the experimental design. E.D., A.T. and C.G. analysed data and conducted the scientific writing. A.T. and C.G. performed the experimental work. A.C. analysed the Affymetrix data. F.O. and F.B. provided molecular biology support. V.P. performed the TOP/FOP assay. M.P., C.D. and S.D. provided scientific input and reagents.
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Supplementary Figures S1, S2, S3, S4, S5, S6, S7 and Supplementary Methods (PDF 1417 kb)
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Taddei, A., Giampietro, C., Conti, A. et al. Endothelial adherens junctions control tight junctions by VE-cadherin-mediated upregulation of claudin-5. Nat Cell Biol 10, 923–934 (2008). https://doi.org/10.1038/ncb1752
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DOI: https://doi.org/10.1038/ncb1752
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