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JXB Advance Access published online on June 20, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri211
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org
Received January 10, 2005
Accepted April 27, 2005

RESEARCH PAPER

Epothilone D affects cell cycle and microtubular pattern in plant cells

Gerd Hause 1, Sandra Lischewski 2, Ludger A. Wessjohann 2, and Bettina Hause 2*

1 Biozentrum der Universität, Martin-Luther-Universität Halle-Wittenberg, Weinbergweg 22, D-06120 Halle (Saale), Germany
2 Leibniz-Institut für Pflanzenbiochemie, Weinberg 3, PO Box 110432, D-06120 Halle (Saale), Germany

* To whom correspondence should be addressed.
Bettina Hause, E-mail: bhause{at}ipb-halle.de


  Abstract

Epothilones, macrocyclic lactones from culture filtrates of the myxobacterium Sorangium cellulosum, are known as taxol-like microtubular drugs in human medicine. To date, nothing is known about the effect of epothilones on microtubules (MTs) in plant cells and/or on the plant cell cycle. As shown in this report, the treatment of tomato cell suspension cultures with epothilone D produced a continuous increase in the mitotic index. Dose-response curves revealed that epothilone D alters the mitotic index at concentrations as low as 1.5 µM. Mitotic arrest was already visible after only 2 h of treatment, and 55% of the cells were arrested after 24 h. As shown by immunocytological methods, abnormal spindles are formed during metaphase, which leads to a random distribution of chromosomes in the whole cell and prevents the formation of a metaphase plate. The process of chromosome decondensation does not seem to be affected, because micronuclei form at the same place with the distributed chromosomes. This suggests that epothilone D influences the stability of plant MTs mainly during metaphase of the mitotic cycle. In metaphase, the effects of epothilone D seem to be irreversible, because cells with an abnormal spindle could not be recovered after removal of the drug.

Keywords: Cell cycle; epothilone; immunofluorescence; Lycopersicon esculentum; microtubuli; mitosis; mitotic arrest.
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