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© The Rockefeller University Press, 0021-9525/1998//147 $5.00
The Journal of Cell Biology, Volume 143, Number 1, , 1998 147-157

Motile Properties of Vimentin Intermediate Filament Networks in Living Cells

Northwestern University Medical School, Department of Cell and Molecular Biology, Chicago, Illinois 60611

The motile properties of intermediate filament (IF) networks have been studied in living cells expressing vimentin tagged with green fluorescent protein (GFP-vimentin). In interphase and mitotic cells, GFP-vimentin is incorporated into the endogenous IF network, and accurately reports the behavior of IF. Time-lapse observations of interphase arrays of vimentin fibrils demonstrate that they are constantly changing their configurations in the absence of alterations in cell shape. Intersecting points of vimentin fibrils, or foci, frequently move towards or away from each other, indicating that the fibrils can lengthen or shorten. Fluorescence recovery after photobleaching shows that bleach zones across fibrils rapidly recover their fluorescence. During this recovery, bleached zones frequently move, indicating translocation of fibrils. Intriguingly, neighboring fibrils within a cell can exhibit different rates and directions of movement, and they often appear to extend or elongate into the peripheral regions of the cytoplasm. In these same regions, short filamentous structures are also seen actively translocating. All of these motile properties require energy, and the majority appear to be mediated by interactions of IF with microtubules and microfilaments.

Key Words: intermediate filaments • vimentin • microtubules • microfilaments • green fluorescent protein

Abbreviations used in this paper: GFP, green fluorescent protein; IF, intermediate filaments; MT, microtubules; MF, microfilaments; FRAP, fluorescence recovery after photobleaching; t_1/2, recovery half-time.

The work has been supported by the National Institute of General Medical Sciences and is in partial fulfillment of the Ph.D degree for M. Yoon.

Address all correspondence to Robert D. Goldman, Northwestern University Medical School, Department of Cell and Molecular Biology, 303 E. Chicago Ave., Chicago, IL 60611. Tel.: 312-503-4215. Fax: 312-503-0954. E-mail: r-goldman{at}nwu.edu

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