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Mechanical coordination in motor ensembles revealed using engineered artificial myosin filaments

Nature Nanotechnology volume 10, pages 696–700 (2015)Cite this article

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Abstract

The sarcomere of muscle is composed of tens of thousands of myosin motors that self-assemble into thick filaments and interact with surrounding actin-based thin filaments in a dense, near-crystalline hexagonal lattice1. Together, these actin–myosin interactions enable large-scale movement and force generation, two primary attributes of muscle. Research on isolated fibres has provided considerable insight into the collective properties of muscle, but how actin–myosin interactions are coordinated in an ensemble remains poorly understood2. Here, we show that artificial myosin filaments, engineered using a DNA nanotube scaffold, provide precise control over motor number, type and spacing. Using both dimeric myosin V- and myosin VI-labelled nanotubes, we find that neither myosin density nor spacing has a significant effect on the gliding speed of actin filaments. This observation supports a simple model of myosin ensembles as energy reservoirs that buffer individual stochastic events to bring about smooth, continuous motion. Furthermore, gliding speed increases with cross-bridge compliance, but is limited by Brownian effects. As a first step to reconstituting muscle motility, we demonstrate human β-cardiac myosin-driven gliding of actin filaments on DNA nanotubes.

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Figure 1: Formation of synthetic myosin filaments using DNA nanotubes.
Figure 2: Density and number of myosins on synthetic filaments have negligible effect on gliding speeds.
Figure 3: Stochastic simulation of actin gliding along myosin filaments predicts actin length-independent gliding speed for sufficiently high motor number.
Figure 4: Actin gliding on synthetic human β-cardiac myosin filaments.

References

  1. Huxley, H. E. The mechanism of muscular contraction. Science 164, 1356–1365 (1969).

    Article  CAS  Google Scholar 

  2. Guérin, T., Prost, J., Martin, P. & Joanny, J. F. Coordination and collective properties of molecular motors: theory. Curr. Opin. Cell Biol. 22, 14–20 (2010).

    Article  Google Scholar 

  3. De La Cruz, E. M. & Ostap, E. M. Relating biochemistry and function in the myosin superfamily. Curr. Opin. Cell Biol. 16, 61–67 (2004).

    Article  CAS  Google Scholar 

  4. Uyeda, T. Q., Kron, S. J. & Spudich, J. A. Myosin step size. Estimation from slow sliding movement of actin over low densities of heavy meromyosin. J. Mol. Biol. 214, 699–710 (1990).

    Article  CAS  Google Scholar 

  5. Sommese, R. F. et al. Molecular consequences of the R453C hypertrophic cardiomyopathy mutation on human β-cardiac myosin motor function. Proc. Natl Acad. Sci. USA 110, 12607–12612 (2013).

    Article  CAS  Google Scholar 

  6. Harris, D. E. & Warshaw, D. M. Smooth and skeletal muscle myosin both exhibit low duty cycles at zero load in vitro. J. Biol. Chem. 268, 14764–14768 (1993).

    CAS  Google Scholar 

  7. Piazzesi, G. et al. Skeletal muscle performance determined by modulation of number of myosin motors rather than motor force or stroke size. Cell 131, 784–795 (2007).

    Article  CAS  Google Scholar 

  8. Spudich, J. A. Hypertrophic and dilated cardiomyopathy: four decades of basic research on muscle lead to potential therapeutic approaches to these devastating genetic diseases. Biophys. J. 106, 1236–1249 (2014).

    Article  CAS  Google Scholar 

  9. Baker, J. E., Brosseau, C., Joel, P. B. & Warshaw, D. M. The biochemical kinetics underlying actin movement generated by one and many skeletal muscle myosin molecules. Biophys. J. 82, 2134–2147 (2002).

    Article  CAS  Google Scholar 

  10. Derr, N. D. et al. Tug-of-war in motor protein ensembles revealed with a programmable DNA origami scaffold. Science 338, 662–665 (2012).

    Article  CAS  Google Scholar 

  11. Hariadi, R. F., Cale, M. & Sivaramakrishnan, S. Myosin lever arm directs collective motion on cellular actin network. Proc. Natl Acad. Sci. USA 111, 4091–4096 (2014).

    Article  CAS  Google Scholar 

  12. Walcott, S., Warshaw, D. M. & Debold, E. P. Mechanical coupling between myosin molecules causes differences between ensemble and single-molecule measurements. Biophys. J. 103, 501–510 (2012).

    Article  CAS  Google Scholar 

  13. Rock, R. S., Rief, M., Mehta, A. D. & Spudich, J. A. In vitro assays of processive myosin motors. Methods 22, 373–381 (2000).

    Article  CAS  Google Scholar 

  14. Lu, H. et al. Collective dynamics of elastically coupled myosin V motors. J. Biol. Chem. 287, 27753–27761 (2012).

    Article  CAS  Google Scholar 

  15. Yin, P. et al. Programming DNA tube circumferences. Science 321, 824–826 (2008).

    Article  CAS  Google Scholar 

  16. Ali, M. Y. et al. Myosin Va and myosin VI coordinate their steps while engaged in an in vitro tug of war during cargo transport. Proc. Natl Acad. Sci. USA 108, E535–E541 (2011).

    Article  CAS  Google Scholar 

  17. Hilbert, L., Cumarasamy, S., Zitouni, N. B., Mackey, M. C. & Lauzon, A. M. The kinetics of mechanically coupled myosins exhibit group size-dependent regimes. Biophys. J. 105, 1466–1474 (2013).

    Article  CAS  Google Scholar 

  18. Rief, M. et al. Myosin-V stepping kinetics: a molecular model for processivity. Proc. Natl Acad. Sci. USA 97, 9482–9486 (2000).

    Article  CAS  Google Scholar 

  19. Rock, R. S. et al. Myosin VI is a processive motor with a large step size. Proc. Natl Acad. Sci. USA 98, 13655–13659 (2001).

    Article  CAS  Google Scholar 

  20. Gebhardt, J. C. M., Clemen, A. E. M., Jaud, J. & Rief, M. Myosin-V is a mechanical ratchet. Proc. Natl Acad. Sci. USA 103, 8680–8685 (2006).

    Article  CAS  Google Scholar 

  21. Veigel, C., Molloy, J. E., Schmitz, S. & Kendrick-Jones, J. Load-dependent kinetics of force production by smooth muscle myosin measured with optical tweezers. Nature Cell Biol. 5, 980–986 (2003).

    Article  CAS  Google Scholar 

  22. Walsh, R., Rutland, C., Thomas, R. & Loughna, S. Cardiomyopathy: a systematic review of disease-causing mutations in myosin heavy chain 7 and their phenotypic manifestations. Cardiology 115, 49–60 (2010).

    Article  CAS  Google Scholar 

  23. Weith, A. et al. Unique single molecule binding of cardiac myosin binding protein-C to actin and phosphorylation-dependent inhibition of actomyosin motility requires 17 amino acids of the motif domain. J. Mol. Cell. Cardiol. 52, 219–227 (2012).

    Article  CAS  Google Scholar 

  24. Resnicow, D. I., Deacon, J. C., Warrick, H. M., Spudich, J. A. & Leinwand, L. A. Functional diversity among a family of human skeletal muscle myosin motors. Proc. Natl Acad. Sci. USA 107, 1053–1058 (2010).

    Article  CAS  Google Scholar 

  25. Trybus, K. M., Freyzon, Y., Faust, L. Z. & Sweeney, H. L. Spare the rod, spoil the regulation: the necessity for a myosin rod. Proc. Natl Acad. Sci. USA 94, 48–52 (1997).

    Article  CAS  Google Scholar 

  26. Huang, J., Nagy, S. S., Koide, A., Rock, R. S. & Koide, S. A peptide tag system for facile purification and single-molecule immobilization. Biochemistry 48, 11834–11836 (2009).

    Article  CAS  Google Scholar 

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Acknowledgements

The authors thank M. Westfall, D. Smith and L. Hilbert for useful discussions. Research was funded by the American Heart Association Scientist Development Grant (13SDG14270009), National Institutes of Health (NIH) grants 1DP2 CA186752-01 and 1-R01-GM-105646-01-A1 to S.S. and NIH grants GM33289 and HL117138 to J.A.S. R.F.S. is a Life Sciences Research Foundation Fellow. R.E.T. is supported by the NIH (F32 HL123247-02) and A.S.A. is supported by a Lucile Packard CHRI Postdoctoral Award.

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Author notes

  1. S. Sivaramakrishnan

    Present address: Present address: Department of Genetics, Cell Biology & Development, College of Biological Sciences, University of Minnesota Twin-Cities, 420 Washington Avenue SE, 4-130 MCB, Minneapolis, Minnesota 55455, USA,

  2. R. F. Hariadi and R. F. Sommese: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, 48109, Michigan, USA

    R. F. Hariadi, R. F. Sommese & S. Sivaramakrishnan

  2. Department of Biochemistry, Stanford University School of Medicine, Stanford, 94305, California, USA

    A. S. Adhikari, R. E. Taylor, S. Sutton & J. A. Spudich

  3. Department of Biophysics, University of Michigan, Ann Arbor, 48109, Michigan, USA

    S. Sivaramakrishnan

  4. Department of Biomedical Engineering, University of Michigan, Ann Arbor, 48109, Michigan, USA

    S. Sivaramakrishnan

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Contributions

R.F.H., R.F.S., A.S.A., R.E.T., J.A.S. and S.S. planned and designed experiments. R.F.H., R.F.S., A.S.A., R.E.T. and S.Su. performed experiments and analysed the results. R.F.H. and S.S. performed the mathematical modelling. R.F.H., R.F.S., J.A.S. and S.S. wrote the manuscript.

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Correspondence to S. Sivaramakrishnan.

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Hariadi, R., Sommese, R., Adhikari, A. et al. Mechanical coordination in motor ensembles revealed using engineered artificial myosin filaments. Nature Nanotech 10, 696–700 (2015). https://doi.org/10.1038/nnano.2015.132

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