Regulation of the actin cycle in vivo by actin filament severing
- James L. McGrath*,†,‡,§,
- Eric A. Osborn*,†,‡,
- Yanik S. Tardy¶,
- C. Forbes Dewey, Jr.†, and
- John H. Hartwig*
- *Hematology Division, Brigham and Women's Hospital, Boston, MA 02115; †Fluid Mechanics Laboratory, Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139; and ¶Biomedical Engineering Laboratory, Swiss Federal Institute of Technology, 1015 Lausanne, Switzerland
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Edited by Thomas P. Stossel, Harvard Medical School, Boston, MA, and approved March 20, 2000 (received for review January 19, 2000)
Abstract
Cycling of actin subunits between monomeric and filamentous phases is essential for cell crawling behavior. We investigated actin filament turnover rates, length, number, barbed end exposure, and binding of cofilin in bovine arterial endothelial cells moving at different speeds depending on their position in a confluent monolayer. Fast-translocating cells near the wound edge have short filament lifetimes compared with turnover values that proportionately increase in slower moving cells situated at increasing distances from the wound border. Contrasted with slow cells exhibiting slow actin filament turnover speeds, fast cells have less polymerized actin, shorter actin filaments, more free barbed ends, and less actin-associated cofilin. Cultured primary fibroblasts manifest identical relationships between speed and actin turnover as the endothelial cells, and fast fibroblasts expressing gelsolin have higher actin turnover rates than slow fibroblasts that lack this actin-severing protein. These results implicate actin filament severing as an important control mechanism for actin cycling in cells.
Footnotes
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↵ ‡ J.L.M. and E.A.O. contributed equally to this work.
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↵ § To whom reprint requests should be addressed at: Johns Hopkins University, Department of Biomedical Engineering, 720 Rutland Avenue, Ross 724, Baltimore, MD 21205-2196. E-mail: jmcgrath{at}bme.jhu.edu.
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This paper was submitted directly (Track II) to the PNAS office.
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Article published online before print: Proc. Natl. Acad. Sci. USA, 10.1073/pnas.100023397.
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Article and publication date are at www.pnas.org/cgi/doi/10.1073/pnas.100023397
- Abbreviations:
- PAF,
- photoactivation fluorescence;
- FRAP,
- fluorescence recovery after photobleaching;
- BAEC,
- bovine aortic endothelial cell;
- ADF,
- actin depolymerizing factor
- Copyright © The National Academy of Sciences
