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ActuAtor, a molecular tool for generating force in living cells: Controlled deformation of intracellular structures

Hideki Nakamura, Elmer Rho, Daqi Deng, Shiva Razavi, Hideaki T. Matsubayashi, View ORCID ProfileTakanari Inoue
doi: https://doi.org/10.1101/2020.03.30.016360
Hideki Nakamura
1Johns Hopkins University School of Medicine, Department of Cell Biology and Center for Cell Dynamics, 855 N. Wolfe Street, Baltimore, MD, 21205, USA
3Kyoto University Graduate School of Engineering, Department of Synthetic Chemistry and Biological Chemistry, Katsura Int’tech Center, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto, 615-8530, Japan
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  • For correspondence: nakamura@sbchem.kyoto-u.ac.jp jctinoue@jhmi.edu
Elmer Rho
1Johns Hopkins University School of Medicine, Department of Cell Biology and Center for Cell Dynamics, 855 N. Wolfe Street, Baltimore, MD, 21205, USA
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Daqi Deng
1Johns Hopkins University School of Medicine, Department of Cell Biology and Center for Cell Dynamics, 855 N. Wolfe Street, Baltimore, MD, 21205, USA
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Shiva Razavi
2Johns Hopkins University School of Medicine, Department of Biomedical Engineering, 855 N. Wolfe Street, Baltimore, MD, 21205, USA
4Massachusetts Institute of Technology, Department of Biomedical Engineering, Cambridge, MA, 02139, USA
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Hideaki T. Matsubayashi
1Johns Hopkins University School of Medicine, Department of Cell Biology and Center for Cell Dynamics, 855 N. Wolfe Street, Baltimore, MD, 21205, USA
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Takanari Inoue
1Johns Hopkins University School of Medicine, Department of Cell Biology and Center for Cell Dynamics, 855 N. Wolfe Street, Baltimore, MD, 21205, USA
2Johns Hopkins University School of Medicine, Department of Biomedical Engineering, 855 N. Wolfe Street, Baltimore, MD, 21205, USA
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  • ORCID record for Takanari Inoue
  • For correspondence: nakamura@sbchem.kyoto-u.ac.jp jctinoue@jhmi.edu
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Summary

Mechanical force underlies fundamental cell functions such as division, migration and differentiation. While physical probes and devices revealed cellular mechano-responses, how force is translated inside cells to exert output functions remains largely unknown, due to the limited techniques to manipulate force intracellularly. By engineering an ActA protein, an actin nucleation promoting factor derived from Listeria monocytogenes, and implementing this in protein dimerization paradigms, we developed a molecular tool termed ActuAtor, with which actin polymerization can be triggered at intended subcellular locations to generate constrictive force in a rapidly inducible manner. The ActuAtor operation led to striking deformation of target intracellular structures including mitochondria, Golgi apparatus, nucleus, and non-membrane-bound RNA granules. Based on functional analysis before and after organelle deformation, we found the form-function relationship of mitochondria to be generally marginal. The modular design and genetically-encoded nature enable wide applications of ActuAtor for studies of intracellular mechanobiology processes.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted March 31, 2020.
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ActuAtor, a molecular tool for generating force in living cells: Controlled deformation of intracellular structures
Hideki Nakamura, Elmer Rho, Daqi Deng, Shiva Razavi, Hideaki T. Matsubayashi, Takanari Inoue
bioRxiv 2020.03.30.016360; doi: https://doi.org/10.1101/2020.03.30.016360
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ActuAtor, a molecular tool for generating force in living cells: Controlled deformation of intracellular structures
Hideki Nakamura, Elmer Rho, Daqi Deng, Shiva Razavi, Hideaki T. Matsubayashi, Takanari Inoue
bioRxiv 2020.03.30.016360; doi: https://doi.org/10.1101/2020.03.30.016360

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