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Feedback regulation through myosin II confers robustness on RhoA signalling at E-cadherin junctions

Nature Cell Biology volume 17, pages 12821293 (2015) | Download Citation

Abstract

Actomyosin at the epithelial zonula adherens (ZA) generates junctional tension for tissue integrity and morphogenesis. This requires the RhoA GTPase, which establishes a strikingly stable active zone at the ZA. Mechanisms must then exist to confer robustness on junctional RhoA signalling at the population level. We now identify a feedback network that generates a stable mesoscopic RhoA zone out of dynamic elements. The key is scaffolding of ROCK1 to the ZA by myosin II. ROCK1 protects junctional RhoA by phosphorylating Rnd3 to prevent the cortical recruitment of the Rho suppressor, p190B RhoGAP. Combining predictive modelling and experimentation, we show that this network constitutes a bistable dynamical system that is realized at the population level of the ZA. Thus, stability of the RhoA zone is an emergent consequence of the network of interactions that allow myosin II to feedback to RhoA.

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Acknowledgements

We thank our laboratory colleagues for their unstinting support and advice during the course of this project, and our colleagues elsewhere for their kind gifts of reagents. We also thank M. Zerial and L. Tucker-Kellogg for thought-provoking discussions, M. Naghibosadat for her help with cloning and E. Moussa for his help in nano-ablation analysis. This work was supported by the National Health and Medical Research Council of Australia (1044041, 1037320, 1067405), The Kids Cancer Project of the Oncology Research Foundation, the EMPathy National Collaborative Research Program (CG-10-04) of the National Breast Cancer Foundation (Australia), a University of Queensland Early Career Research Grant to G.A.G. (2012003354) and an ANZ Trustees PhD Scholarship in Medical Research to R.P.

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Author notes

    • Guillermo A. Gomez
    •  & Alpha S. Yap

    These authors contributed equally to this work.

Affiliations

  1. Division of Cell Biology and Molecular Medicine, Institute for Molecular Bioscience, The University of Queensland, St Lucia, Brisbane, Queensland 4072, Australia

    • Rashmi Priya
    • , Guillermo A. Gomez
    • , Srikanth Budnar
    • , Suzie Verma
    • , Hayley L. Cox
    • , Nicholas A. Hamilton
    •  & Alpha S. Yap

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Contributions

R.P., G.A.G. and A.S.Y. conceived the project and designed the experiments. R.P. performed most of the experiments in collaboration with G.A.G. for nano-ablation, FRAP and Rho biosensor experiments, with assistance from S.V. for western blotting and immunoprecipitation experiments. G.A.G. and N.A.H. performed the mathematical modelling. H.L.C. performed experiments. S.B. performed the cloning and characterization of AHPH mutants. R.P. and G.A.G. analysed the data. R.P., G.A.G. and A.S.Y. wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Guillermo A. Gomez or Alpha S. Yap.

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Videos

  1. 1.

    GFP-AHPH localizes at the zonula adherens of epithelial cells.

    Z-stacks of GFP-AHPH and RFP-UtrCH acquired by spinning disc confocal microscopy.

  2. 2.

    GFP-AHPH exhibits stability on the time scale of minutes.

    Time-lapse imaging of GFP-APHPH (transfected in MCF-7 cells) acquired over a span of 30 min.

  3. 3.

    ROCK-1 inhibition causes accumulation of p190B Rho GAP at the cell–cell junctions.

    MCF-7 cells were transfected with GFP-p190B RhoGAP and time-lapsed imaging was performed briefly before and after addition of Y-27632 (30 μM) (20-min post-treatment).

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DOI

https://doi.org/10.1038/ncb3239

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