Abstract

Multicellularity in animals requires dynamic maintenance of cell–cell contacts. Intercellularly ligated cadherins recruit numerous proteins to form supramolecular complexes that connect with the actin cytoskeleton and support force transmission. However, the molecular organization within such structures remains unknown. Here we mapped protein organization in cadherin-based adhesions by super-resolution microscopy, revealing a multi-compartment nanoscale architecture, with the plasma-membrane-proximal cadherin–catenin compartment segregated from the actin cytoskeletal compartment, bridged by an interface zone containing vinculin. Vinculin position is determined by α-catenin, and following activation, vinculin can extend 30 nm to bridge the cadherin–catenin and actin compartments, while modulating the nanoscale positions of the actin regulators zyxin and VASP. Vinculin conformational activation requires tension and tyrosine phosphorylation, regulated by Abl kinase and PTP1B phosphatase. Such modular architecture provides a structural framework for mechanical and biochemical signal integration by vinculin, which may differentially engage cadherin–catenin complexes with the actomyosin machinery to regulate cell adhesions.

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Acknowledgements

We thank C. Ajo-Franklin (Lawrence Berkeley National Laboratory), G. Shtengel and H. Hess (Howard Hughes Medical Institute, Janelia Research Campus) for generous help with super-resolution microscopy instrumentation and analysis. We thank H. Chen (Protein Expression Facility, Mechanobiology Institute, Singapore) for generation of mutant constructs, H. T. Ong and A. Sathe (Mechanobiology Institute) for assistance with FRET analysis, C. Zhang (Mechanobiology Institute) for assistance with 3D graphical model, and J. Yan (National University of Singapore) for critical reading of the manuscript. P.K., C.B., Y.Wang, A.R., T.S. and R.Z.-B. are supported by Singapore National Research Foundation Fellowship awarded to P.K. (NRF-NRFF-2011-04), R.Z.-B. (NRF-RF2009-RF001-074), and Competitive Research Programme (NRF2012NRF-CRP001-084) to P.K. and R.Z.-B. Y.T. is supported by Mechanobiology Institute and National University of Singapore Startup Grants, and a Singapore Ministry of Education Tier 2 grant (MOE2015-T2-1-116). B.L. is supported by the European Research Council under the European Union’s Seventh Framework Programme (FP7/2007–2013/ERC grant agreement no. 617233), the Mechanobiology Institute, and the Institut Universitaire de France, and USPC-NUS funding. B.L. and R.-M.M. are also supported by CNRS, the Human Frontier Science Program (grant RGP0040/2012) and Agence Nationale de la Recherche (ANR 2010 Blan1515).

Author information

Author notes

    • Yilin Wang
    • , Talgat Sailov
    •  & Michelle A. Baird

    Present addresses: Department of Biology, South University of Science and Technology, Shenzhen 518055, China (Y.W.); Singapore Centre on Environmental Life Sciences Engineering, Nanyang Technological University, Singapore 637551, Republic of Singapore (T.S.); National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA (M.A.B.).

    • Michael W. Davidson

    Deceased.

Affiliations

  1. Mechanobiology Institute, Singapore 117411, Republic of Singapore

    • Cristina Bertocchi
    • , Yilin Wang
    • , Andrea Ravasio
    • , Yusuke Hara
    • , Yao Wu
    • , Talgat Sailov
    • , Ronen Zaidel-Bar
    • , Yusuke Toyama
    • , Benoit Ladoux
    •  & Pakorn Kanchanawong
  2. National High Magnetic Field Laboratory, The Florida State University, Tallahassee, Florida 32310, USA

    • Michelle A. Baird
    •  & Michael W. Davidson
  3. Department of Biological Science, The Florida State University, Tallahassee, Florida 32306, USA

    • Michael W. Davidson
  4. Department of Biomedical Engineering, National University of Singapore, Singapore 117583, Republic of Singapore

    • Ronen Zaidel-Bar
    •  & Pakorn Kanchanawong
  5. Department of Biological Sciences, National University of Singapore, Singapore 117543, Republic of Singapore

    • Yusuke Toyama
  6. Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604, Republic of Singapore

    • Yusuke Toyama
  7. Institut Jacques Monod, Université Paris Diderot, CNRS UMR 7592, Paris, France

    • Benoit Ladoux
    •  & Rene-Marc Mege

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Contributions

C.B. and Y.Wang performed the super-resolution imaging experiments and conducted data analysis. C.B. and A.R. performed and analysed FRET experiments. C.B., A.R., Y.H. and Y.T. designed and C.B. performed and analysed laser ablation experiments. Y.Wu and R.Z.-B. performed imaging of Eph4 cell–cell junctions by astigmatism-based 3D super-resolution microscopy. C.B., T.S., M.B., M.W.D., B.L. and R.-M.M. designed and generated fusion constructs, and provided new reagents and analytical tools. C.B. and P.K. designed the study and wrote the manuscript. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Pakorn Kanchanawong.

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https://doi.org/10.1038/ncb3456

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