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Microscopic analysis of polymerization dynamics with individual actin filaments

Nature Cell Biology volume 4, pages 666–673 (2002)Cite this article

Abstract

The polymerization–depolymerization dynamics of actin is a key process in a variety of cellular functions. Many spectroscopic studies have been performed in solution, but studies on single actin filaments have just begun. Here, we show that the time course of polymerization of individual filaments consists of a polymerization phase and a subsequent steady-state phase. During the steady-state phase, a treadmilling process of elongation at the barbed end and shortening at the pointed end occurs, in which both components of the process proceed at approximately the same rate. The time correlation of length fluctuation of the filaments in the steady-state phase showed that the polymerization–depolymerization dynamics follow a diffusion (stochastic) process, which cannot be explained by simple association and dissociation of monomers at both ends of the filaments.

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Figure 1: Fluorescence micrographs of actin(Ca) polymerization.
Figure 2: Time course of actin(Ca) polymerization at various actin concentrations.
Figure 3: The effects of G-actin(Ca) concentration on filament growth during polymerization.
Figure 4: Frequency of actin(Ca) filament length fluctuation in a steady-state phase.
Figure 5: Analysis of the polymerization dynamics of actin(Mg).
Figure 6: Square of the width of the length fluctuation histogram versus correlation time.

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Acknowledgements

This research was partly supported by Grants-in-Aid for Scientific Research, Scientific Research on Priority Areas and Bio-venture Projects from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and by Grants-in-Aid from the Japan Science and Technology Corporation (CREST) and the Mitsubishi Foundation. We thank W. Rozycki for reading the manuscript. We also thank colleagues at Waseda University and CREST for their encouragement and support. I.F. is a research fellow of the Japan Society for the Promotion of Science.

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  1. Department of Physics, School of Science and Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, 169-8555, Tokyo, Japan

    Ikuko Fujiwara, Shin Takahashi, Hisashi Tadakuma, Takashi Funatsu & Shin'ichi Ishiwata

  2. Advanced Research Institute for Science and Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, 169-8555, Tokyo, Japan

    Shin'ichi Ishiwata

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Correspondence to Shin'ichi Ishiwata.

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Fujiwara, I., Takahashi, S., Tadakuma, H. et al. Microscopic analysis of polymerization dynamics with individual actin filaments. Nat Cell Biol 4, 666–673 (2002). https://doi.org/10.1038/ncb841

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