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The effect of diffusion, depolymerization and nucleation promoting factors on actin gel growth

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Abstract

In eukaryotic cells, localized actin polymerization is able to deform the plasma membrane and push the cell forward. Depolymerization of actin filaments and diffusion of actin monomers ensure the availability of monomers at sites of polymerization, and therefore these processes must play an active role in cellular actin dynamics. Here we reveal experimental evidence that actin gel growth can be limited by monomer diffusion, consistent with theoretical predictions. We study actin gels formed on beads coated with ActA (and ActA fragments), the bacterial factor responsible for actin-based movement of Listeria monocytogenes. We observe a saturation of gel thickness with increasing bead radius, the signature of diffusion control. Data analysis using an elastic model of actin gel growth gives an estimate of 2×10−8 cm−2 s−1 for the diffusion coefficient of actin monomers through the gel, ten times less than in buffer, and in agreement with literature values in bulk cytoskeleton, providing corroboration of our model. The depolymerization rate of actin filaments and the elastic modulus of the gel are also evaluated. Furthermore, we qualitatively examine the different actin gels produced when ActA fragments interact with either VASP or the Arp2/3 complex.

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Acknowledgements

This work was supported by Curie and CNRS fellowships (J.P.). We thank Vincent Noireaux for performing the treadmilling experiment shown in Fig. 3. We thank Marie-France Carlier for the gift of Arp2/3 and VASP, and Henry Higgs for the p41-Arc antibody.

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  1. Laboratoire Physicochimie “Curie”, UMR168 Institut Curie/CNRS, 11 rue Pierre et Marie Curie, 75231 , Paris cedex 05, France

    Julie Plastino, Ioannis Lelidis, Jacques Prost & Cécile Sykes

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  1. Julie Plastino
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  2. Ioannis Lelidis
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  3. Jacques Prost
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  4. Cécile Sykes
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Correspondence to Cécile Sykes.

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Plastino, J., Lelidis, I., Prost, J. et al. The effect of diffusion, depolymerization and nucleation promoting factors on actin gel growth. Eur Biophys J 33, 310–320 (2004). https://doi.org/10.1007/s00249-003-0370-3

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