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Cofilin promotes rapid actin filament turnover in vivo

Nature volume 388, pages 78–82 (1997)Cite this article

Abstract

The ability of actin filaments to function in cell morphogenesis and motility is coupled to their capacity for rapid assembly and disassembly. Because disassembly in vitro is much slower than in vivo, cellular factors that stimulate disassembly have long been assumed to exist. Although numerous proteins can affect actin dynamics in vitro, demonstration of in vivo relevance of these effects has not been achieved. We have used genetics and an actin-inhibitor in yeast to demonstrate that rapid cycles of actin assembly and disassembly depend on the small actin-binding protein cofilin, and that cofilin stimulates filament disassembly. These results may explain why cofilin is ubiquitous in eukaryotes and is essential for viability in every organism in which its function has been tested genetically. Magnitudes of disassembly defects in cofilin mutants in vivo were found to be correlated closely with the magnitudes of disassembly defects observed in vitro, supporting our conclusions. Furthermore, these cofilin mutants provided an opportunity to distinguish in living cells those actin functions that depend specifically on filament turnover (endocytosis) from those that do not (cortical actin patch motility).

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Figure 1: Cells were grown at 20 °C to A600 = 0.3, shifted to 34 °C for 3 h and fixed with 5% formaldehyde.
Figure 2: Filamentous actin was visualized by rhodamine–phalloidin staining in wild-type (a–c), cof1–5 (d–f) and cof1–22 (g–i) strains at 0, 2 and 10 min after the addition of 400 µM Lat-A to the cultures at 25 °C.
Figure 3: The effects of cofilin mutations on actin-filament depolymerization in vivo and in vitro.
Figure 4: Effects of cofilin mutants on endocytosis and actin patch movement.

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Acknowledgements

We thank K. Ayscough for the Lat-A disassembly procedure; L. Belmont, B. Goode and K. Kozminski for comments on the manuscript; and A. Mallavarapu and T. Mitchison for help in recording patch movements. This work was supported by long-term fellowships from the European Molecular Biology Organization and Human Frontier Science Program (to P.L.) and by grants from the NIH and American Cancer Society (to D.G.D.).

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  1. Department of Molecular and Cell Biology, 401 Barker Hall, University of California, Berkeley, 94720, California, USA

    Pekka Lappalainen & David G. Drubin

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  1. Pekka Lappalainen
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Correspondence to David G. Drubin.

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Lappalainen, P., Drubin, D. Cofilin promotes rapid actin filament turnover in vivo. Nature 388, 78–82 (1997). https://doi.org/10.1038/40418

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