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Vol. 15, Issue 3, 1374-1386, March 2004

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-Tubulin Plays an Essential Role in the Coordination of Mitotic Events

Natalie L. Prigozhina , C. Elizabeth Oakley, Amanda M. Lewis, Tania Nayak, Stephen A. Osmani, and Berl R. Oakley 

Department of Molecular Genetics, The Ohio State University, Columbus, Ohio 43210

Submitted June 16, 2003; Revised November 11, 2003; Accepted November 12, 2003
Monitoring Editor: Frank Solomon

Recent data from multiple organisms indicate that -tubulin has essential, but incompletely defined, functions in addition to nucleating microtubule assembly. To investigate these functions, we examined the phenotype of mipAD159, a cold-sensitive allele of the -tubulin gene of Aspergillus nidulans. Immunofluorescence microscopy of synchronized material revealed that at a restrictive temperature mipAD159 does not inhibit mitotic spindle formation. Anaphase A was inhibited in many nuclei, however, and after a slight delay in mitosis (6% of the cell cycle period), most nuclei reentered interphase without dividing. In vivo observations of chromosomes at a restrictive temperature revealed that mipAD159 caused a failure of the coordination of late mitotic events (anaphase A, anaphase B, and chromosomal disjunction) and nuclei reentered interphase quickly even though mitosis was not completed successfully. Time-lapse microscopy also revealed that transient mitotic spindle abnormalities, in particular bent spindles, were more prevalent in mipAD159 strains than in controls. In experiments in which microtubules were depolymerized with benomyl, mipAD159 nuclei exited mitosis significantly more quickly (as judged by chromosomal condensation) than nuclei in a control strain. These data reveal that -tubulin has an essential role in the coordination of late mitotic events, and a microtubule-independent function in mitotic checkpoint control.

Online version of this article contains supplementary video material for some figures. Online version is available at www.molbiolcell.org.

Present address: Department of Cell Biology, MB-39, Room MBB201, The Scripps Research Institute, 10550 North Torrey Pines Rd., La Jolla, CA 92037.

Corresponding author. E-mail address: oakley.2{at}osu.edu.

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