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Published 8 May 2006. doi:10.1083/jcb.200511124
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 173, Number 3, 341-348
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Focal adhesions are hotspots for keratin filament precursor formation



Reinhard Windoffer, Anne Kölsch, Stefan Wöll, and Rudolf E. Leube

Department of Anatomy and Cell Biology, Johannes Gutenberg University, 55128 Mainz, Germany

Correspondence to Rudolf E. Leube: leube{at}uni-mainz.de

Recent studies showed that keratin filament (KF) formation originates primarily from sites close to the actin-rich cell cortex. To further characterize these sites, we performed multicolor fluorescence imaging of living cells and found drastically increased KF assembly in regions of elevated actin turnover, i.e., in lamellipodia. Abundant KF precursors (KFPs) appeared within these areas at the distal tips of actin stress fibers, moving alongside the stress fibers until their integration into the peripheral KF network. The earliest KFPs were detected next to actin-anchoring focal adhesions (FAs) and were only seen after the establishment of FAs in emerging lamellipodia. Tight spatiotemporal coupling of FAs and KFP formation were not restricted to epithelial cells, but also occurred in nonepithelial cells and cells producing mutant keratins. Finally, interference with FA formation by talin short hairpin RNA led to KFP depletion. Collectively, our results support a major regulatory function of FAs for KF assembly, thereby providing the basis for coordinated shaping of the entire cytoskeleton during cell relocation and rearrangement.

R. Windoffer and A. Kölsch contributed equally to this paper.

Abbreviations used in this paper: ECFP, enhanced CFP; EYFP, enhanced YFP; IF, intermediate filament; FA, focal adhesion; HK, human keratin; KF, keratin filament; KFP, KF precursor; RFP, red fluorescent protein; shRNA, short hairpin RNA.


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