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Science 4 April 2003:
Vol. 300. no. 5616, pp. 142 - 145
DOI: 10.1126/science.1082026
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Reports

Rapid Actin Transport During Cell Protrusion

Daniel Zicha,1 Ian M. Dobbie,2 Mark R. Holt,3 James Monypenny,1 Daniel Y. H. Soong,3 Colin Gray,1 Graham A. Dunn3*

Transformed rat fibroblasts expressing two variants of green fluorescent protein, each fused to beta -actin, were used to study actin dynamics during cell protrusion. The recently developed FLAP (fluorescence localization after photobleaching) method permits the tracking of one fluorophore after localized photobleaching by using the other as a colocalized reference. Here, by visualizing the ratio of bleached to total molecules, we found that actin was delivered to protruding zones of the leading edge of the cell at speeds that exceeded 5 micrometers per second. Monte Carlo modeling confirmed that this flow cannot be explained by diffusion and may involve active transport.

1 Light Microscopy, Cancer Research UK, Lincoln's Inn Fields Laboratories, London WC2A 3PX, UK.
2 Genome Stability Laboratory, Department of Biochemistry, National University of Ireland, Galway, Ireland.
3 The Randall Centre, New Hunt's House, Guy's Campus, King's College London, London SE1 1UL, UK.
*   To whom correspondence should be addressed. E-mail: graham.a.dunn{at}kcl.ac.uk

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