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Vol. 14, Issue 2, 642-657, February 2003
*Max-Planck-Institut für Terrestrische
Mikrobiologie, Karl-von-Frisch-Stra Growth of most eukaryotic cells requires directed transport along
microtubules (MTs) that are nucleated at nuclear-associated microtubule
organizing centers (MTOCs), such as the centrosome and the fungal
spindle pole body (SPB). Herein, we show that the pathogenic fungus
Ustilago maydis uses different MT nucleation sites to
rearrange MTs during the cell cycle. In vivo observation of green
fluorescent protein-MTs and MT plus-ends, tagged by a fluorescent EB1
homologue, provided evidence for antipolar MT orientation and dispersed
cytoplasmic MT nucleating centers in unbudded cells. On budding
e, D-35043 Marburg, Germany;
Departments of Molecular Genetics and Plant
Biology, The Ohio State University, Columbus, Ohio 43210; and
§Department of Food Microbiology, School of
Medicine, University of Tokushima, Tokushima 770-8503, Japan
-tubulin containing MTOCs formed at the bud neck, and MTs
reorganized with >85% of all minus-ends being focused toward the
growth region. Experimentally induced lateral budding resulted in MTs
that curved out of the bud, again supporting the notion that polar
growth requires polar MT nucleation. Depletion or overexpression of
Tub2, the -tubulin from U. maydis, affected MT number
in interphase cells. The SPB was inactive in G2 phase but continuously
recruited -tubulin until it started to nucleate mitotic MTs. Taken
together, our data suggest that MT reorganization in U.
maydis depends on cell cycle-specific nucleation at dispersed
cytoplasmic sites, at a polar MTOC and the SPB.
Present address: Institut für Medizinische
Mikrobiologie, Immunologie und Hygiene, Trogerstrae 4a, D-81675
München, Germany.
Corresponding author. E-mail address:
gero.steinberg{at}staff.uni-marburg.de.
Online version of this
article contains video material for some figures. Online version
available at www.molbiolcell.org.
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