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ROCK and Dia have opposing effects on adherens junctions downstream of Rho

Nature Cell Biology volume 4, pages 408–415 (2002)Cite this article

Abstract

Adherens junctions (AJs) are crucial for maintaining the integrity of epithelial tissues and are often disrupted during tumour progression. Rho family proteins have been shown to regulate adherens junctions. We find that activation of the effector kinase ROCK and acto-myosin contraction disrupts AJs downstream of Rho. In contrast, signalling through the Rho effector Dia1 is required to ensure a dynamically stable interface between cells and the maintenance of adherens junction complexes. The ability of Dia1 to regulate the actin network is crucial for the localization of adherens junction components to the cell periphery.

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Figure 1: Effect of Rho inhibition on cell junctions.
Figure 2: Rho is required for the stability of the cell periphery.
Figure 3: Dia1 functions downstream from Rho to maintain adherens junction complexes.
Figure 4: Overexpression of RhoC is more effective than RhoA at disrupting adherens junctions.
Figure 5: ROCK and acto-myosin contractility are required downstream of RhoC to disrupt adherens junctions.
Figure 6: Y-27632 promotes adherens junction formation in SW620 colon carcinoma cells.
Figure 7: Opposing pathways downstream of Rho regulating adherens junctions.

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References

  1. Yap, A. S., Brieher, W. M. & Gumbiner, B. M. Molecular and functional analysis of cadherin-based adherens junctions. Annu. Rev. Cell Dev. Biol. 13, 119–146 (1997).

    Article  CAS  Google Scholar 

  2. Fukata, M. & Kaibuchi, K. Rho-family GTPases in cadherin-mediated cell–cell adhesion. Nature Rev. Mol. Cell Biol. 2, 887–897 (2001).

    Article  CAS  Google Scholar 

  3. Van Aelst, L. & D'Souza-Schorey, C. Rho GTPases and signaling networks. Genes Dev. 11, 2295–2322 (1997).

    Article  CAS  Google Scholar 

  4. Ridley, A. J. Rho GTPases and cell migration. J. Cell Sci. 114, 2713–2722 (2001).

    CAS  Google Scholar 

  5. Braga, V. M., Machesky, L. M., Hall, A. & Hotchin, N. A. The small GTPases Rho and Rac are required for the establishment of cadherin-dependent cell–cell contacts. J. Cell Biol. 137, 1421–1431 (1997).

    Article  CAS  Google Scholar 

  6. Takaishi, K., Sasaki, T., Kotani, H., Nishioka, H. & Takai, Y. Regulation of cell–cell adhesion by rac and rho small G proteins in MDCK cells. J. Cell Biol. 139, 1047–1059 (1997).

    Article  CAS  Google Scholar 

  7. Jou, T. S. & Nelson, W. J. Effects of regulated expression of mutant RhoA and Rac1 small GTPases on the development of epithelial (MDCK) cell polarity. J. Cell Biol. 142, 85–100 (1998).

    Article  CAS  Google Scholar 

  8. Bishop, A. L. & Hall, A. Rho GTPases and their effector proteins. Biochem. J. 348, 241–255 (2000).

    Article  CAS  Google Scholar 

  9. Kimura, K. et al. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase). Science 273, 245–248 (1996).

    Article  CAS  Google Scholar 

  10. Maekawa, M. et al. Signaling from Rho to the actin cytoskeleton through protein kinases ROCK and LIM-kinase. Science 285, 895–898 (1999).

    Article  CAS  Google Scholar 

  11. Vincent, S. & Settleman, J. The PRK2 kinase is a potential effector target of both Rho and Rac GTPases and regulates actin cytoskeletal organization. Mol. Cell. Biol. 17, 2247–2256 (1997).

    Article  CAS  Google Scholar 

  12. Watanabe, N. et al. p140mDia, a mammalian homolog of Drosophila diaphanous, is a target protein for Rho small GTPase and is a ligand for profilin. EMBO J. 16, 3044–3056 (1997).

    Article  CAS  Google Scholar 

  13. Alberts, A. S. Identification of a carboxyl-terminal diaphanous-related formin homology protein autoregulatory domain. J. Biol. Chem. 276, 2824–2830 (2001).

    Article  CAS  Google Scholar 

  14. Ishizaki, T. et al. Coordination of microtubules and the actin cytoskeleton by the Rho effector mDia1. Nature Cell Biol. 3, 8–14 (2001).

    Article  CAS  Google Scholar 

  15. Palazzo, A. F., Cook, T. A., Alberts, A. S. & Gundersen, G. G. mDia mediates Rho-regulated formation and orientation of stable microtubules. Nature Cell Biol. 3, 723–729 (2001).

    Article  CAS  Google Scholar 

  16. Bhowmick, N. A. et al. Transforming Growth Factor-β1 Mediates Epithelial to Mesenchymal Transdifferentiation through a RhoA-dependent Mechanism. Mol. Biol. Cell 12, 27–36 (2001).

    Article  CAS  Google Scholar 

  17. Wojciak-Stothard, B., Potempa, S., Eichholtz, T. & Ridley, A. J. Rho and Rac but not Cdc42 regulate endothelial cell permeability. J. Cell Sci. 114, 1343–1355 (2001).

    CAS  Google Scholar 

  18. Calautti, E. et al. Fyn tyrosine kinase is a downstream mediator of Rho/PRK2 function in keratinocyte cell–cell adhesion. J. Cell Biol. 156, 137–148 (2002).

    Article  CAS  Google Scholar 

  19. Hiscox, S. & Jiang, W. G. Expression of E-cadherin, α, β and γ-catenin in human colorectal cancer. Anticancer Res. 17, 1349–1354 (1997).

    CAS  Google Scholar 

  20. Sahai, E. & Marshall, C. J. RhoGTPases and Cancer. Nature Reviews Cancer 2, 133–142 (2002).

    Article  Google Scholar 

  21. Gayet, J. et al. Extensive characterization of genetic alterations in a series of human colorectal cancer cell lines. Oncogene 20, 5025–5032 (2001).

    Article  CAS  Google Scholar 

  22. Fukata, M. et al. Involvement of IQGAP1, an effector of Rac1 and Cdc42 GTPases, in cell–cell dissociation during cell scattering. Mol. Cell Biol. 21, 2165–2183 (2001).

    Article  CAS  Google Scholar 

  23. Sahai, E., Alberts, A. S. & Treisman, R. RhoA effector mutants reveal distinct effector pathways for cytoskeletal reorganization, SRF activation and transformation. EMBO J. 17, 1350–1361 (1998).

    Article  CAS  Google Scholar 

  24. Davies, S. P., Reddy, H., Caivano, M. & Cohen, P. Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem J. 351, 95–105 (2000).

    Article  CAS  Google Scholar 

  25. Sahai, E., Olson, M. F. & Marshall, C. J. Cross-talk between Ras and Rho signalling pathways in transformation favours proliferation and increased motility. EMBO J. 20, 755–766 (2001).

    Article  CAS  Google Scholar 

  26. Noren, N. K., Niessen, C. M., Gumbiner, B. M. & Burridge, K. Cadherin engagement regulates Rho family GTPases. J. Biol. Chem. 276, 33305–33308 (2001).

    Article  CAS  Google Scholar 

  27. Quinlan, M. P. & Hyatt, J. L. Establishment of the circumferential actin filament network is a prerequisite for localization of the cadherin–catenin complex in epithelial cells. Cell Growth Differ. 10, 839–854 (1999).

    CAS  Google Scholar 

  28. Vasioukhin, V., Bauer, C., Yin, M. & Fuchs, E. Directed actin polymerization is the driving force for epithelial cell–cell adhesion. Cell 100, 209–219 (2000).

    Article  CAS  Google Scholar 

  29. Quinlan, M. P. Rac regulates the stability of the adherens junction and its components, thus affecting epithelial cell differentiation and transformation. Oncogene 18, 6434–6442 (1999).

    Article  CAS  Google Scholar 

  30. Tokman, M. G., Porter, R. A. & Williams, C. L. Regulation of cadherin-mediated adhesion by the small GTP-binding protein Rho in small cell lung carcinoma cells. Cancer Res. 57, 1785–1793 (1997).

    CAS  Google Scholar 

  31. Hill, C. S., Wynne, J. & Treisman, R. The Rho family GTPases RhoA, Rac1, and CDC42Hs regulate transcriptional activation by SRF. Cell 81, 1159–1170 (1995).

    Article  CAS  Google Scholar 

  32. Tominaga, T. et al. Diaphanous-related formins bridge Rho GTPase and Src tyrosine kinase signaling. Mol. Cell 5, 13–25 (2000).

    Article  CAS  Google Scholar 

  33. Ishizaki, T. et al. p160ROCK, a Rho-associated coiled-coil forming protein kinase, works downstream of Rho and induces focal adhesions. FEBS Lett. 404, 118–124 (1997).

    Article  CAS  Google Scholar 

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Acknowledgements

E.S. and C.J.M. are funded by Cancer Research UK; C.J.M. is a Gibb Life Fellow of Cancer Research UK. We thank M. Fukata for GFP–α-catenin, A. Alberts for GFP–mDia1ΔN, H. Paterson for help with microscopy, D. Croft and M. Olson for comments on the manuscript.

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Authors and Affiliations

  1. Cancer Research UK Centre for Cell and Molecular Biology, Institute of Cancer Research, 237 Fulham Road, London, SW3 6JB, UK

    Erik Sahai & Christopher J. Marshall

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Corresponding author

Correspondence to Christopher J. Marshall.

Supplementary information

Supplementary figure

Figure S1. Effects of Dia mutants on actin and microtubule networks a, HCT116 cells in 1% DCS were injected with expression constructs for GFP-mDia1ΔN, GFPmDia1ΔNK>A or mDia1ΔNΔC. 16 hours after injection the cells were fixed and stained for GFP or myc-epitope to detect exogenous Dia expression, β-tubulin, and F-actin. b, HEK 293 cells were transfected with either mDia1ΔNΔC, RhoAV14 or mDia1ΔNΔC and RhoAV14, 24 hours after transfection cells were fixed and stained for myc-epitope to detect transfected cells and F-actin. (PDF 153 kb)

Supplemetary movie

Movie 1. HCT116 cells in 10% DCS expressing GFP-α-catenin, frames are 60 seconds apart. (AVI 1280 kb)

Supplemetary movie

Movie 2. HCT116 cells in 10% DCS expressing GFP-α-catenin and C3, frames are 60 seconds apart. (AVI 3784 kb)

Supplemetary movie

Movie 3. HCT15 cells in 10% DCS expressing GFP-α-catenin, frames are 60 seconds apart. (AVI 800 kb)

Supplemetary movie

Movie 4. HCT15 cells in 10% DCS expressing GFP-α-catenin and mDia1ΔNΔC, frames are 60 seconds apart. (AVI 2868 kb)

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Sahai, E., Marshall, C. ROCK and Dia have opposing effects on adherens junctions downstream of Rho. Nat Cell Biol 4, 408–415 (2002). https://doi.org/10.1038/ncb796

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