Vascular Endothelial-Cadherin Regulates Cytoskeletal Tension, Cell Spreading, and Focal Adhesions by Stimulating RhoA
10.1091/mbc.e03-10-0745
Crossref journal-article
American Society for Cell Biology (ASCB)
Molecular Biology of the Cell (1076)
Abstract

Changes in vascular endothelial (VE)-cadherin–mediated cell-cell adhesion and integrin-mediated cell-matrix adhesion coordinate to affect the physical and mechanical rearrangements of the endothelium, although the mechanisms for such cross talk remain undefined. Herein, we describe the regulation of focal adhesion formation and cytoskeletal tension by intercellular VE-cadherin engagement, and the molecular mechanism by which this occurs. Increasing the density of endothelial cells to increase cell-cell contact decreased focal adhesions by decreasing cell spreading. This contact inhibition of cell spreading was blocked by disrupting VE-cadherin engagement with an adenovirus encoding dominant negative VE-cadherin. When changes in cell spreading were prevented by culturing cells on a micropatterned substrate, VE-cadherin–mediated cell-cell contact paradoxically increased focal adhesion formation. We show that VE-cadherin engagement mediates each of these effects by inducing both a transient and sustained activation of RhoA. Both the increase and decrease in cell-matrix adhesion were blocked by disrupting intracellular tension and signaling through the Rho-ROCK pathway. In all, these findings demonstrate that VE-cadherin signals through RhoA and the actin cytoskeleton to cross talk with cell-matrix adhesion and thereby define a novel pathway by which cell-cell contact alters the global mechanical and functional state of cells.

Bibliography

Nelson, C. M., Pirone, D. M., Tan, J. L., & Chen, C. S. (2004). Vascular Endothelial-Cadherin Regulates Cytoskeletal Tension, Cell Spreading, and Focal Adhesions by Stimulating RhoA. Molecular Biology of the Cell, 15(6), 2943–2953.

Authors 4
  1. Celeste M. Nelson (first)
  2. Dana M. Pirone (additional)
  3. John L. Tan (additional)
  4. Christopher S. Chen (additional)
References 65 Referenced 152
  1. Abercrombie, M., and Turner, A.A. (1978). Contract reactions influencing cell locomotion of a mouse sarcoma in culture.Med. Biol.56, 299-303.
  2. Alexander, J.S., and Elrod, J.W. (2002). Extracellular matrix, junctional integrity and matrix metalloproteinase interactions in endothelial permeability regulation.J. Anat.200, 561-574. (10.1046/j.1469-7580.2002.00057.x)
  3. Amano, M., Fukata, Y., and Kaibuchi, K. (2000). Regulation and functions of Rho-associated kinase.Exp. Cell Res.261, 44-51. (10.1006/excr.2000.5046)
  4. Anastasiadis, P.Z., and Reynolds, A.B. (2001). Regulation of Rho GTPases by p120-catenin.Curr. Opin. Cell Biol.13, 604-610. (10.1016/S0955-0674(00)00258-1)
  5. Arregui, C., Pathre, P., Lilien, J., and Balsamo, J. (2000). The nonreceptor tyrosine kinase fer mediates cross-talk between N-cadherin and beta1-integrins.J. Cell Biol.149, 1263-1274. (10.1083/jcb.149.6.1263)
  6. Assoian, R.K., and Schwartz, M.A. (2001). Coordinate signaling by integrins and receptor tyrosine kinases in the regulation of G1 phase cell-cycle progression.Curr. Opin. Genet. Dev.11, 48-53. (10.1016/S0959-437X(00)00155-6)
  7. Balaban, N.Q.et al. (2001). Force and focal adhesion assembly: a close relationship studied using elastomeric micropatterned substrates.Nat. Cell Biol.3, 466-472. (10.1038/35074532)
  8. Ballestrem, C., Hinz, B., Imhof, B.A., and Wehrle-Haller, B. (2001). Marching at the front and dragging behind: differential alphavbeta3-integrin turnover regulates focal adhesion behavior.J. Cell Biol.155, 1319-1332. (10.1083/jcb.200107107)
  9. Betson, M., Lozano, E., Zhang, J., and Braga, V.M.M. (2002). Rac activation upon cell-cell contact formation is dependent on signaling from the epidermal growth factor receptor.J. Biol. Chem.277, 36962-36969. (10.1074/jbc.M207358200)
  10. Bishop, A.L., and Hall, A. (2000). Rho GTPases and their effector proteins.Biochem. J.348, 241-255. (10.1042/bj3480241)
  11. Braga, V.M. (2002). Cell-cell adhesion and signalling.Curr. Opin. Cell Biol.14, 546-556. (10.1016/S0955-0674(02)00373-3)
  12. Calautti, E., Grossi, M., Mammucari, C., Aoyama, Y., Pirro, M., Ono, Y., Li, J., and Dotto, G.P. (2002). Fyn tyrosine kinase is a downstream mediator of Rho/PRK2 function in keratinocyte cell-cell adhesion.J. Cell Biol.156, 137-148. (10.1083/jcb.200105140)
  13. Carmeliet, P.et al. (1999). Targeted deficiency or cytosolic truncation of the VE-cadherin gene in mice impairs VEGF-mediated endothelial survival and angiogenesis.Cell98, 147-157. (10.1016/S0092-8674(00)81010-7)
  14. Charrasse, S., Meriane, M., Comunale, F., Blangy, A., and Gauthier-Rouviere, C. (2002). N-cadherin-dependent cell-cell contact regulates Rho GTPases and beta-catenin localization in mouse C2C12 myoblasts.J. Cell Biol.158, 953-965. (10.1083/jcb.200202034)
  15. Chen, C.S., Alonso, J.L., Ostuni, E., Whitesides, G.M., and Ingber, D.E. (2003). Cell shape provides global control of focal adhesion assembly.Biochem. Biophys. Res. Commun.307, 355-361. (10.1016/S0006-291X(03)01165-3)
  16. Chrzanowska-Wodnicka, M., and Burridge, K. (1996). Rho-stimulated contractility drives the formation of stress fibers and focal adhesions.J. Cell Biol.133, 1403-1415. (10.1083/jcb.133.6.1403)
  17. Clark, E.A., King, W.G., Brugge, J.S., Symons, M., and Hynes, R.O. (1998). Integrin-mediated signals regulated by members of the Rho family of GT-Pases.J. Cell Biol.142, 573-586. (10.1083/jcb.142.2.573)
  18. Davies, P.F.et al. (1997). Spatial relationships in early signaling events of flow-mediated endothelial mechanotransduction.Annu. Rev. Physiol.59, 527-549. (10.1146/annurev.physiol.59.1.527)
  19. Dejana, E., Bazzoni, G., and Lampugnani, M.G. (1999). Vascular endothelial (VE)-cadherin: only an intercellular glue?Exp. Cell Res.252, 13-19. (10.1006/excr.1999.4601)
  20. Dudek, S.M., and Garcia, J.G. (2001). Cytoskeletal regulation of pulmonary vascular permeability.J. Appl. Physiol.91, 1487-1500. (10.1152/jappl.2001.91.4.1487)
  21. Etienne-Manneville, S., and Hall, A. (2002). Rho GTPases in cell biology.Nature420, 629-635. (10.1038/nature01148)
  22. Ferrara, N., Gerber, H.P., and LeCouter, J. (2003). The biology of VEGF and its receptors.Nat. Med.9, 669-676. (10.1038/nm0603-669)
  23. Geiger, B., and Bershadsky, A. (2001). Assembly and mechanosensory function of focal contacts.Curr. Opin. Cell Biol.13, 584-592. (10.1016/S0955-0674(00)00255-6)
  24. Geiger, B., and Bershadsky, A. (2002). Exploring the neighborhood: adhesion-coupled cell mechanosensors.Cell110, 139-142. (10.1016/S0092-8674(02)00831-0)
  25. Grosheva, I., Shtutman, M., Elbaum, M., and Bershadsky, A.D. (2001). p120 catenin affects cell motility via modulation of activity of Rho-family GTPases: a link between cell-cell contact formation and regulation of cell locomotion.J. Cell Sci.114, 695-707. (10.1242/jcs.114.4.695)
  26. Gumbiner, B.M. (1996). Cell adhesion: the molecular basis of tissue architecture and morphogenesis.Cell84, 345-357. (10.1016/S0092-8674(00)81279-9)
  27. Hirose, M., Ishizaki, T., Watanabe, N., Uehata, M., Kranenburg, O., Moolenaar, W.H., Matsamura, F., Maekawa, M., Bito, H., and Narumiya, S. (1998). Molecular dissection of the Rho-associated protein kinase (p160ROCK)-regulated neurite remodeling in neuroblastoma N1E-115 cells.J. Cell Biol.141, 1625-1636. (10.1083/jcb.141.7.1625)
  28. Hodivala, K.J., and Watt, F.M. (1994). Evidence that cadherins play a role in the downregulation of integrin expression that occurs during keratinocyte terminal differentiation.J. Cell Biol.124, 589-600. (10.1083/jcb.124.4.589)
  29. Hood, J.D., and Cheresh, D.A. (2002). Role of integrins in cell invasion and migration.Nat. Rev. Cancer2, 91-100. (10.1038/nrc727)
  30. Hormia, M., Badley, R.A., Lehto, V.P., and Virtanen, I. (1985). Actomyosin organization in stationary and migrating sheets of cultured human endothelial cells.Exp. Cell Res.157, 116-126. (10.1016/0014-4827(85)90156-9)
  31. Jalink, K., van Corven, E.J., Hengeveld, T., Morii, N., Narumiya, S., and Moolenaar, W.H. (1994). Inhibition of lysophosphatidate- and thrombin-induced neurite retraction and neuronal cell rounding by ADP ribosylation of the small GTP-binding protein Rho.J. Cell Biol.126, 801-810. (10.1083/jcb.126.3.801)
  32. Juliano, R.L. (2002). Signal transduction by cell adhesion receptors and the cytoskeleton: functions of integrins, cadherins, selectins, and immunoglobulin-superfamily members.Annu. Rev. Pharmacol. Toxicol.42, 283-323. (10.1146/annurev.pharmtox.42.090401.151133)
  33. Ko, K.S., Arora, P.D., and McCulloch, C.A. (2001). Cadherins mediate intercellular mechanical signaling in fibroblasts by activation of stretch-sensitive calcium-permeable channels.J. Biol. Chem.276, 35967-35977. (10.1074/jbc.M104106200)
  34. Kouklis, P., Konstantoulaki, M., and Malik, A.B. (2003). VE-cadherin-induced Cdc42 signaling regulates formation of membrane protrusions in endothelial cells.J. Biol. Chem.278, 16230-16236. (10.1074/jbc.M212591200)
  35. 10.1091/mbc.01-07-0368
  36. Levenberg, S., Katz, B.Z., Yamada, K.M., and Geiger, B. (1998). Long-range and selective autoregulation of cell-cell or cell-matrix adhesions by cadherin or integrin ligands.J. Cell Sci.111, 347-357. (10.1242/jcs.111.3.347)
  37. Lewis, J.E., Jensen, P.J., Johnson, K.R., and Wheelock, M.J. (1994). E-cadherin mediates adherens junction organization through protein kinase C.J. Cell Sci.107, 3615-3621. (10.1242/jcs.107.12.3615)
  38. Magie, C.R., Pinto-Santini, D., and Parkhurst, S.M. (2002). Rho1 interacts with p120ctn and alpha-catenin, and regulates cadherin-based adherens junction components inDrosophila.Development129, 3771-3782. (10.1242/dev.129.16.3771)
  39. Nakagawa, M., Fukata, M., Yamaga, M., Itoh, N., and Kaibuchi, K. (2001). Recruitment and activation of Rac1 by the formation of E-cadherin-mediated cell-cell adhesion sites.J. Cell Sci.114, 1829-1838. (10.1242/jcs.114.10.1829)
  40. Navarro, P., Caveda, L., Breviario, F., Mandoteanu, I., Lampugnani, M.G., and Dejana, E. (1995). Catenin-dependent and -independent functions of vascular endothelial cadherin.J. Biol. Chem.270, 30965-30972. (10.1074/jbc.270.52.30965)
  41. Nelson, C.M., and Chen, C.S. (2002). Cell-cell signaling by direct contact increases cell proliferation via a PI3K-dependent mechanism.FEBS Lett.514, 238-242. (10.1016/S0014-5793(02)02370-0)
  42. Nelson, C.M., and Chen, C.S. (2003). VE-cadherin simultaneously stimulates and inhibits cell proliferation by altering cytoskeletal structure and tension.J. Cell Sci.116, 3571-3581. (10.1242/jcs.00680)
  43. Nobes, C.D., and Hall, A. (1995). Rho, Rac and Cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia.Cell81, 53-62. (10.1016/0092-8674(95)90370-4)
  44. Noren, N.K., Liu, B.P., Burridge, K., and Kreft, B. (2000). p120 catenin regulates the actin cytoskeleton via rho family GTPases.J. Cell Biol.150, 567-579. (10.1083/jcb.150.3.567)
  45. Noren, N.K., Niessen, C.M., Gumbiner, B.M., and Burridge, K. (2001). Cadherin engagement regulates Rho family GTPases.J. Biol. Chem.276, 33305-33308. (10.1074/jbc.C100306200)
  46. Palecek, S.P., Horwitz, A.F., and Lauffenburger, D.A. (1999). Kinetic model for integrin-mediated adhesion release during cell migration.Ann. Biomed. Eng.27, 219-235. (10.1114/1.176)
  47. Pece, S., and Gutkind, A.S. (2000). Signaling from E-cadherins to the MAPK pathway by the recruitment and activation of epidermal growth factor receptors upon cell-cell contact formation.J. Biol. Chem.275, 41227-41233. (10.1074/jbc.M006578200)
  48. Plopper, G., and Ingber, D.E. (1993). Rapid induction and isolation of focal adhesion complexes.Biochem. Biophys. Res. Commun.193, 571-578. (10.1006/bbrc.1993.1662)
  49. Pokutta, S., and Weis, W.I. (2002). The cytoplasmic face of cell contact sites.Curr. Opin. Sruct. Biol.12, 255-262. (10.1016/S0959-440X(02)00318-4)
  50. Putnam, A.J., Cunningham, J.J., Pillemer, B.B., and Mooney, D.J. (2003). External mechanical strain regulates membrane targeting of Rho GTPases by controlling microtubule assembly.Am. J. Physiol..284, C627-C639. (10.1152/ajpcell.00137.2002)
  51. Ren, X.D., Kiosses, W.B., and Schwartz, M.A. (1999). Regulation of the small GTP-binding protein Rho by cell adhesion and the cytoskeleton.EMBO J.18, 578-585. (10.1093/emboj/18.3.578)
  52. Ren, X.D., Kiosses, W.B., Sieg, D.J., Otey, C.A., Schlaepfer, D.D., and Schwartz, M.A. (2000). Focal adhesion kinase suppresses Rho activity to promote focal adhesion turnover.J. Cell Sci.113, 3673-3678. (10.1242/jcs.113.20.3673)
  53. Ridley, A.J., Paterson, H.F., Johnston, C.L., Diekmann, D., and Hall, A. (1992). The small GTP-binding protein Rac regulates growth factor-induced membrane ruffling.Cell70, 401-410. (10.1016/0092-8674(92)90164-8)
  54. Riveline, D., Zamir, E., Balaban, N.Q., Schwartz, U.S., Ishizaki, T., Narumiya, S., Kam, Z., Geiger, B., and Bershadsky, A. (2001). Focal contacts as mechanosensors: externally applied local force induces growth of focal contacts by an mDia1-dependent and ROCK-independent mechanism.J. Cell Biol.153, 1175-1183. (10.1083/jcb.153.6.1175)
  55. Ryan, P.L., Foty, R.A., Kohn, J., and Steinberg, M.S. (2001). Tissue spreading on implantable substrates is a competitive outcome of cell-cell vs. cell-substratum adhesivity.Proc. Natl. Acad. Sci. USA98, 4323-4327. (10.1073/pnas.071615398)
  56. Savagner, P. (2001). Leaving the neighborhood: molecular mechanisms involved during epithelial-mesenchymal transition.Bioessays23, 912-923. (10.1002/bies.1132)
  57. Shay-Salit, A., Shushy, M., Wolfovitz, E., Yahav, H., Breviario, F., Dejana, E., and Resnick, N. (2002). VEGF receptor 2 and the adherens junction as a mechanical transducer in vascular endothelial cells.Proc. Natl. Acad. Sci. USA99, 9462-9467. (10.1073/pnas.142224299)
  58. Stupack, D.G., and Cheresh, D.A. (2002). Get a ligand, get a life: integrins, signaling and cell survival.J. Cell Sci.115, 3729-3738. (10.1242/jcs.00071)
  59. Vaezi, A., Bauer, C., Vasioukhin, V., and Fuchs, E. (2002). Actin cable dynamics and Rho/Rock orchestrate a polarized cytoskeletal architecture in the early steps of assembling a stratified epithelium.Dev. Cell3, 367-381. (10.1016/S1534-5807(02)00259-9)
  60. van Leeuwen, F.N., van Delft, S., Kain, H.E., van der Kammen, R.A., and Collard, J.G. (1999). Rac regulates phosphorylation of the myosin-II heavy chain, actinomyosin disassembly and cell spreading.Nat. Cell Biol.1, 242-248. (10.1038/12068)
  61. Vestweber, D. (2000). Molecular mechanisms that control endothelial cell contacts.J. Pathol.190, 281-291. (10.1002/(SICI)1096-9896(200002)190:3<281::AID-PATH527>3.0.CO;2-Z)
  62. Webb, D.J., Parsons, J.T., and Horwitz, A.F. (2002). Adhesion assembly, disassembly and turnover in migrating cells–over and over and over again.Nat. Cell Biol.4, E97-E100. (10.1038/ncb0402-e97)
  63. Yap, A.S., and Kovacs, E.M. (2003). Direct cadherin-activated cell signaling: a view from the plasma membrane.J. Cell Biol.160, 11-16. (10.1083/jcb.200208156)
  64. Yap, A.S., and Manley, S.W. (1993). Contact inhibition of cell spreading: a mechanism for the maintenance of thyroid cell aggregation in vitro.Exp. Cell Res.208, 121-127. (10.1006/excr.1993.1229)
  65. Zhu, A.J., and Watt, F.M. (1996). Expression of a dominant negative cadherin mutant inhibits proliferation and stimulates terminal differentiation of human epidermal keratinocytes.J. Cell Sci.109, 3013-3023. (10.1242/jcs.109.13.3013)
Dates
Type When
Created 21 years, 4 months ago (April 12, 2004, 8:53 p.m.)
Deposited 4 years, 2 months ago (June 15, 2021, 10:45 a.m.)
Indexed 2 weeks, 6 days ago (Aug. 5, 2025, 8:41 a.m.)
Issued 21 years, 2 months ago (June 1, 2004)
Published 21 years, 2 months ago (June 1, 2004)
Published Print 21 years, 2 months ago (June 1, 2004)
Funders 0

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@article{Nelson_2004, title={Vascular Endothelial-Cadherin Regulates Cytoskeletal Tension, Cell Spreading, and Focal Adhesions by Stimulating RhoA}, volume={15}, ISSN={1939-4586}, url={http://dx.doi.org/10.1091/mbc.e03-10-0745}, DOI={10.1091/mbc.e03-10-0745}, number={6}, journal={Molecular Biology of the Cell}, publisher={American Society for Cell Biology (ASCB)}, author={Nelson, Celeste M. and Pirone, Dana M. and Tan, John L. and Chen, Christopher S.}, year={2004}, month=jun, pages={2943–2953} }