Abstract
Directional polymerization of actin filaments in branched networks is one of the most powerful force-generating systems in eukaryotic cells1. Growth of densely cross-linked actin networks drives cell crawling2, intracellular transport of vesicles and organelles3,4, and movement of intracellular pathogens such as Listeria monocytogenes5. Using a modified atomic force microscope (AFM), we obtained force–velocity (Fv) measurements of growing actin networks in vitro until network elongation ceased at the stall force. We found that the growth velocity of a branched actin network against increasing forces is load-independent over a wide range of forces before a convex decline to stall. Surprisingly, when force was decreased on a growing network, the velocity increased to a value greater than the previous velocity, such that two or more stable growth velocities can exist at a single load. These results demonstrate that a single Fv relationship does not capture the complete behaviour of this system, unlike other molecular motors in cells, because the growth velocity depends on loading history rather than solely on the instantaneous load.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Pollard, T. D. & Borisy, G. G. Cellular motility driven by assembly and disassembly of actin filaments. Cell 113, 549–549 (2003).
Theriot, J. A. & Mitchison, T. J. Actin microfilament dynamics in locomoting cells. Nature 352, 126–131 (1991).
Rozelle, A. L. et al. Phosphatidylinositol 4,5-bisphosphate induces actin-based movement of raft-enriched vesicles through WASP-Arp2/3. Curr. Biol. 10, 311–320 (2000).
Merrifield, C. J. et al. Endocytic vesicles move at the tips of actin tails in cultured mast cells. Nature Cell Biol. 1, 72–74 (1999).
Theriot, J. A., Mitchison, T. J., Tilney, L. G. & Portnoy, D. A. The rate of actin-based motility of intracellular Listeria-Monocytogenes equals the rate of actin polymerization. Nature 357, 257–260 (1992).
Cameron, L. A., Footer, M. J., van Oudenaarden, A. & Theriot, J. A. Motility of ActA protein-coated microspheres driven by actin polymerization. Proc. Natl Acad. Sci. USA 96, 4908–4913 (1999).
Loisel, T. P., Boujemaa, R., Pantaloni, D. & Carlier, M. F. Reconstitution of actin-based motility of Listeria and Shigella using pure proteins. Nature 401, 613–616 (1999).
Upadhyaya, A. & van Oudenaarden, A. Biomimetic systems for studying actin-based motility. Curr. Biol. 13, R734–R744 (2003).
Bustamante, C., Chemla, Y. R., Forde, N. R. & Izhaky, D. Mechanical processes in biochemistry. Annu. Rev. Biochem. 73, 705–748 (2004).
Dogterom, M. & Yurke, B. Measurement of the force–velocity relation for growing microtubules. Science 278, 856–860 (1997).
Mogilner, A. & Oster, G. Force generation by actin polymerization II: The elastic ratchet and tethered filaments. Biophys. J. 84, 1591–1605 (2003).
Mogilner, A. & Oster, G. Cell motility driven by actin polymerization. Biophys. J. 71, 3030–3045 (1996).
Gerbal, F., Chaikin, P., Rabin, Y. & Prost, J. An elastic analysis of Listeria monocytogenes propulsion. Biophys. J. 79, 2259–2275 (2000).
Carlsson, A. E. Growth velocities of branched actin networks. Biophys. J. 84, 2907–2918 (2003).
Dickinson, R. B. & Purich, D. L. Clamped-filament elongation model for actin-based motors. Biophys. J. 82, 605–617 (2002).
McGrath, J. L. et al. The force–velocity relationship for the actin-based motility of Listeria monocytogenes. Curr. Biol. 13, 329–332 (2003).
Wiesner, S. et al. A biomimetic motility assay provides insight into the mechanism of actin-based motility. J. Cell Biol. 160, 387–398 (2003).
Marcy, Y., Prost, J., Carlier, M. F. & Sykes, C. Forces generated during actin-based propulsion: A direct measurement by micromanipulation. Proc. Natl Acad. Sci. USA 101, 5992–5997 (2004).
Cameron, L. A., Giardini, P. A., Soo, F. S. & Theriot, J. A. Secrets of actin-based motility revealed by a bacterial pathogen. Nature Rev. Mol. Cell Biol. 1, 110–119 (2000).
Schwartz, I. M., Ehrenberg, M., Bindschadler, M. & McGrath, J. L. The role of substrate curvature in actin-based pushing forces. Curr. Biol. 14, 1094–1098 (2004).
Wang, M. D. et al. Force and velocity measured for single molecules of RNA polymerase. Science 282, 902–907 (1998).
Smith, D. E. et al. The bacteriophage phi 29 portal motor can package DNA against a large internal force. Nature 413, 748–752 (2001).
Finer, J. T., Simmons, R. M. & Spudich, J. A. Single myosin molecule mechanics — piconewton forces and nanometer steps. Nature 368, 113–119 (1994).
Hill, T. L. & Kirschner, M. W. Bioenergetics and kinetics of microtubule and actin filament assembly-disassembly. Int. Rev. Cytol. 78, 1–125 (1982).
Spudich, J. A. & Watt, S. Regulation of rabbit skeletal muscle contraction.1. Biochemical studies of interaction of Tropomyosin–Troponin complex with actin and proteolytic fragments of Myosin. J. Biol. Chem. 246, 4866–4871 (1971).
Kellogg, D. R., Mitchison, T. J. & Alberts, B. M. Behavior of microtubules and actin-filaments in living Drosophila embryos. Development 103, 675–686 (1988).
Hutter, J. L. & Bechhoefer, J. Calibration of atomic-force microscope tips. Rev. Sci. Instrum. 64, 1868–1873 (1993).
Acknowledgements
We thank J. W. Shaevitz, A. P. Liu, and J. L. Choy for helpful discussions and careful reading of the manuscript, as well as the entire Fletcher laboratory for support. We are also grateful to R. L. Jeng, C. Le Clainche, and M. J. Footer for assistance in protein preparation. This work was supported by a National Defense Science and Engineering Graduate Fellowship to O.C. and an National Science Foundation Career Award to D.A.F.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Information
Supplementary Information and Reference plus Supplementary figures S1, S2 and S3 (PDF 672 kb)
Rights and permissions
About this article
Cite this article
Parekh, S., Chaudhuri, O., Theriot, J. et al. Loading history determines the velocity of actin-network growth. Nat Cell Biol 7, 1219–1223 (2005). https://doi.org/10.1038/ncb1336
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ncb1336
This article is cited by
-
Mechanisms and roles of podosomes and invadopodia
Nature Reviews Molecular Cell Biology (2023)
-
A computational modeling of invadopodia protrusion into an extracellular matrix fiber network
Scientific Reports (2022)
-
On the Stability of Surface Growth: The Effect of a Compliant Surrounding Medium
Journal of Elasticity (2022)
-
Stress fiber growth and remodeling determines cellular morphomechanics under uniaxial cyclic stretch
Biomechanics and Modeling in Mechanobiology (2022)
-
Forces generated by lamellipodial actin filament elongation regulate the WAVE complex during cell migration
Nature Cell Biology (2021)