Myosin II tension sensors visualize force generation within the actin cytoskeleton in living cells
Nonmuscle myosin II (NMII) generates cytoskeletal forces that drive cell division, embryogenesis, muscle contraction and many other cellular functions. However, at present there is no method that can directly measure the forces generated by myosins in living cells. Here, we describe a Förster resonance energy transfer (FRET)-based tension sensor that can detect myosin-associated force along the filamentous actin network. Fluorescence lifetime imaging microscopy (FLIM)-FRET measurements indicate that the forces generated by NMII isoform B (NMIIB) exhibit significant spatial and temporal heterogeneity as a function of donor lifetime and fluorophore energy exchange. These measurements provide a proxy for inferred forces that vary widely along the actin cytoskeleton. This initial report highlights the potential utility of myosin-based tension sensors in elucidating the roles of cytoskeletal contractility in a wide variety of contexts.
Authors
- Divya Kota
- Diego Ramallo
- Ryan G. Hart (ORCID: https://orcid.org/0000-0002-1208-6652)
- Mark O. Huising (ORCID: https://orcid.org/0000-0002-6594-2205)
- Alexander R. Dunn (ORCID: https://orcid.org/0000-0001-6096-4600)
- Karla Otterpohl
- Fangjia Li (ORCID: https://orcid.org/0009-0003-2430-0665)
- Indra Chandrasekar (ORCID: https://orcid.org/0000-0003-3822-0570)
- Andrew Price (ORCID: https://orcid.org/0000-0002-1782-7587)
- Jing Liu (ORCID: https://orcid.org/0000-0002-4912-4560)
- Mengdi Zhang (ORCID: https://orcid.org/0000-0003-4423-4899)
- Steve Smith
Institutions
- University of Indianapolis (US)
- South Dakota School of Mines and Technology (US)
- University of South Dakota (US)
- Purdue University West Lafayette (US)
- University of Sioux Falls (US)
- Sanford Research (US)
- Indiana University – Purdue University Indianapolis (US)
- University of California, Davis (US)
- Stanford University (US)
Publication Details
- Journal
- Journal of Cell Science
- Published
- 2024-10-06
- DOI
- https://doi.org/10.1242/jcs.262281
- Citations
- 7
- Primary Topic
- Cardiomyopathy and Myosin Studies
- Type
- article
- Field-Weighted Citation Impact
- 2.41
Funders
- National Science Foundation
- National Institutes of Health
- National Institute of Diabetes and Digestive and Kidney Diseases
- Office of Experimental Program to Stimulate Competitive Research