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Activation of a Bacterial Mechanosensitive Channel in Mammalian Cells by Cytoskeletal Stress

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Abstract

Cells can sense a myriad of mechanical stimuli. Mechanosensitive channel of large conductance (MscL) found in bacteria is a well-characterized mechanosensitive channel that rapidly responds to an increase in turgor pressure. Functional expression of MscL in mammalian cells has recently been demonstrated, revealing that molecular delivery or transport can be achieved by charge-induced activation of MscL. Despite a well-accepted mechanism for MscL activation by membrane tension in bacteria, it is not clear whether and how MscL can be opened by other modes of force transduction in mammalian cells. In this work, we used a variety of techniques to characterize the gating of MscL expressed in mammalian cells, using both wild type and a G22S mutant which activates at a lower threshold. In particular, employing a recently developed technique, acoustic tweezing cytometry (ATC), we show that ultrasound actuation of integrin-bound microbubbles can lead to MscL opening and that ATC induced MscL activation was dependent on the functional linkage of the microbubbles with an intact actin cytoskeleton. Our results indicate that localized mechanical stress can mediate opening of MscL that requires force transduction through the actin cytoskeleton, revealing a new mode of MscL activation that may prove to be a useful tool for mechanobiology and drug delivery research.

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Acknowledgments

We would like to thank Dr. Boris Martinac for the E. coli MscL constructs used in this study, and his very thoughtful feedback on our manuscript. We thank the Liu lab for helpful discussions and Dr. Jianping Fu for use of his lab for some experiments in the study. J.H. was supported by the NIH’s Microfluidics in Biomedical Sciences Training Program: NIH NIBIB T32 EB005582. A.P.L. and V.L.M. were supported by the NIH Director’s New Innovator Award: NIH DP2 HL117748-01. C.X.D. and D.C. were supported by funding from the Department of Biomedical Engineering at the University of Michigan. The authors declare no conflict of interests.

Conflict of interest

J. Heureaux, D. Chen, V.L. Murray, C.X. Deng, and A.P. Liu declare that they have no conflicts of interests.

Ethical Standards

No human studies were carried out by the authors for this article. No animal studies were carried out by the authors for this article.

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Authors and Affiliations

  1. Department of Mechanical Engineering, University of Michigan, 2350 Hayward Street, Ann Arbor, MI, 48105, USA

    Johanna Heureaux, Victoria L. Murray & Allen P. Liu

  2. Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA

    Di Chen, Cheri X. Deng & Allen P. Liu

  3. Cellular and Molecular Biology Program, University of Michigan, Ann Arbor, MI, USA

    Allen P. Liu

  4. Biophysics Program, University of Michigan, Ann Arbor, MI, USA

    Allen P. Liu

Authors
  1. Johanna Heureaux
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Correspondence to Allen P. Liu.

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Associate Editor Cynthia A. Reinhart-King oversaw the review of this article.

This paper is part of the 2014 Young Innovators Issue.

Allen P. Liu received a B.Sc. degree in Biochemistry (Honours) from the University of British Columbia, Vancouver, Canada, in 2001. He obtained his Ph.D. in Biophysics in 2007 from the University of California-Berkeley, receiving two fellowships from the Natural Sciences and Engineering Research Council of Canada. As a graduate student in Daniel Fletcher’s lab, he studied the dynamic interplay between actin networks and membrane using a reconstituted system. From 2007 to 2011, he was a post-doctoral fellow in Sandra Schmid and Gaudenz Danuser’s labs in the Department of Cell Biology at The Scripps Research Institute in La Jolla, CA, while holding a post-doctoral fellowship from the Leukemia and Lymphoma Society. By combining live cell total internal reflection fluorescence microscopy and high-content image analysis, he studied the role of cortical tension and clustering of cargo molecules in regulating the dynamics of clathrin-coated pits. Since January 2012, he has been an Assistant Professor in the Department of Mechanical Engineering and Biomedical Engineering at the University of Michigan. He is a recipient of the NIH Director’s New Innovator Award. His current research interests include cell mechanics, mechanobiology, and synthetic biology.

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Heureaux, J., Chen, D., Murray, V.L. et al. Activation of a Bacterial Mechanosensitive Channel in Mammalian Cells by Cytoskeletal Stress. Cel. Mol. Bioeng. 7, 307–319 (2014). https://doi.org/10.1007/s12195-014-0337-8

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