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Published online 2 June 2003. doi:10.1083/jcb.200301102
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© The Rockefeller University Press, 0021-9525/2003/6/853 $5.00
The Journal of Cell Biology, Volume 161, Number 5, 853-859

Report

 Nanometer targeting of microtubules to focal adhesions

Olga Krylyshkina1, Kurt I. Anderson2, Irina Kaverina1, Irene Upmann3, Dietmar J. Manstein3, J. Victor Small1 and Derek K. Toomre4

1 Institute of Molecular Biology, Salzburg A5020, Austria
2 Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany
3 Biophysics Department, Max Planck Institute for Medical Research, D-69120 Heidelberg, Germany
4 Department of Cell Biology, Ludwig Institute for Cancer Research, Yale University School of Medicine, New Haven, CT 06520

Address correspondence to Derek K. Toomre, Dept. of Cell Biology, Ludwig Institute for Cancer Research, Yale University School of Medicine, 333 Cedar Street, PO Box 208002, New Haven, CT 06520. Tel.: (203) 785-4319. Fax: (203) 785-3559. E-mail: derek.toomre{at}yale.edu; or J.V. Small, Austrian Academy of Sciences, Institute of Molecular Biology, Dept. of Cell Biology, Billrothstrasse 11, Salzburg A5020, Austria. Tel.: 43-662-63961-11. Fax: 43-662-63961-40. E-mail: jvsmall{at}imb.oeaw.ac.at

Although cell movement is driven by actin, polarization and directional locomotion require an intact microtubule cytoskeleton that influences polarization by modulating substrate adhesion via specific targeting interactions with adhesion complexes. The fidelity of adhesion site targeting is precise; using total internal reflection fluorescence microscopy (TIRFM), we now show microtubule ends (visualized by incorporation of GFP tubulin) are within 50 nm of the substrate when polymerizing toward the cell periphery, but not when shrinking from it. Multiple microtubules sometimes followed similar tracks, suggesting guidance along a common cytoskeletal element. Use of TIRFM with GFP- or DsRed-zyxin in combination with either GFP-tubulin or GFP–CLIP-170 further revealed that the polymerizing microtubule plus ends that tracked close to the dorsal surface consistently targeted substrate adhesion complexes. This supports a central role for the microtubule tip complex in the guidance of microtubules into adhesion foci, and provides evidence for an intimate cross-talk between microtubule tips and substrate adhesions in the range of molecular dimensions.

Key Words: cytoskeleton; CLIP-170; TIRFM; zyxin; cross-talk

The online version of this article includes supplemental material.

I. Upmann and D.J. Manstein's present address is Institute for Biophysical Chemistry, OE 4350, and Central Unit for Biochemical and Biophysical Systems, OE 8830, Medizinische Hochschule Hannover, D-30623 Hannover, Germany.

* Abbreviation used in this paper: TIRFM, total internal reflection fluorescence microscopy.


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