ActuAtor, a Listeria-inspired molecular tool for physical manipulation of intracellular organizations through de novo actin polymerization

Hideki Nakamura; Elmer Rho; Christopher T Lee; Kie Itoh; Daqi Deng; Satoshi Watanabe; Shiva Razavi; Hideaki T Matsubayashi; Cuncheng Zhu; Eleanor Jung; Padmini Rangamani; Shigeki Watanabe; Takanari Inoue

doi:10.1016/j.celrep.2023.113089

ActuAtor, a Listeria-inspired molecular tool for physical manipulation of intracellular organizations through de novo actin polymerization

Cell Rep. 2023 Oct 31;42(10):113089. doi: 10.1016/j.celrep.2023.113089. Epub 2023 Sep 20.

Authors

Affiliations

¹ Department of Cell Biology and Center for Cell Dynamics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Kyoto University Hakubi Center for Advanced Research, Yoshida-honmachi, Sakyo-ku, Kyoto 606-8501, Japan; Kyoto 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. Electronic address: nakamura@sbchem.kyoto-u.ac.jp.
² Department of Cell Biology and Center for Cell Dynamics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
³ Department of Mechanical and Aerospace Engineering, University of California San Diego, La Jolla, CA 92093, USA.
⁴ Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
⁵ Department of Cell Biology and Center for Cell Dynamics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: jctinoue@jhmi.edu.

Abstract

Form and function are often interdependent throughout biology. Inside cells, mitochondria have particularly attracted attention since both their morphology and functionality are altered under pathophysiological conditions. However, directly assessing their causal relationship has been beyond reach due to the limitations of manipulating mitochondrial morphology in a physiologically relevant manner. By engineering a bacterial actin regulator, ActA, we developed tools termed "ActuAtor" that inducibly trigger actin polymerization at arbitrary subcellular locations. The ActuAtor-mediated actin polymerization drives striking deformation and/or movement of target organelles, including mitochondria, Golgi apparatus, and nucleus. Notably, ActuAtor operation also disperses non-membrane-bound entities such as stress granules. We then implemented ActuAtor in functional assays, uncovering the physically fragmented mitochondria being slightly more susceptible to degradation, while none of the organelle functions tested are morphology dependent. The modular and genetically encoded features of ActuAtor should enable its application in studies of the form-function interplay in various intracellular contexts.

Keywords: CP: Cell biology; actin polymerization; form-function interplay; mitochondria; molecular actuators; physical manipulation; stress granules.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Actins / metabolism
Bacterial Proteins / metabolism
Listeria monocytogenes* / physiology
Listeria* / metabolism
Organelles / metabolism
Polymerization

Substances

Actins
Bacterial Proteins

Abstract

Publication types

MeSH terms

Substances

Grants and funding