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Originally published as MBC in Press, 10.1091/mbc.E05-12-1118 on May 3, 2006

Vol. 17, Issue 7, 3242-3253, July 2006

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Dynein-dependent Motility of Microtubules and Nucleation Sites Supports Polarization of the Tubulin Array in the Fungus Ustilago maydisFormula

Gero Fink, and Gero Steinberg

Max-Planck-Institut für terrestrische Mikrobiologie, D-35043 Marburg, Germany

Submitted December 8, 2005; Revised April 21, 2006; Accepted April 24, 2006
Monitoring Editor: Yixian Zheng

Microtubules (MTs) are often organized by a nucleus-associated MT organizing center (MTOC). In addition, in neurons and epithelial cells, motor-based transport of assembled MTs determines the polarity of the MT array. Here, we show that MT motility participates in MT organization in the fungus Ustilago maydis. In budding cells, most MTs are nucleated by three to six small and motile {gamma}-tubulin–containing MTOCs at the boundary of mother and daughter cell, which results in a polarized MT array. In addition, free MTs and MTOCs move rapidly throughout the cytoplasm. Disruption of MTs with benomyl and subsequent washout led to an equal distribution of the MTOC and random formation of highly motile and randomly oriented MTs throughout the cytoplasm. Within 3 min after washout, MTOCs returned to the neck region and the polarized MT array was reestablished. MT motility and polarity of the MT array was lost in dynein mutants, indicating that dynein-based transport of MTs and MTOCs polarizes the MT cytoskeleton. Observation of green fluorescent protein-tagged dynein indicated that this is achieved by off-loading dynein from the plus-ends of motile MTs. We propose that MT organization in U. maydis involves dynein-mediated motility of MTs and nucleation sites.

This was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E05-12-1118) on May 3, 2006.

Formula The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

Address correspondence to: Gero Steinberg ( gero.steinberg{at}staff.uni-marburg.de)

Abbreviations used: GFP, green fluorescent protein; LatA, latrunculin A; MT, microtubule; MTOC, microtubule organizing center; RFP, red fluorescent protein; YFP, yellow fluorescent protein.




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