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Originally published as MBC in Press, 10.1091/mbc.E04-06-0486 on September 15, 2004

Vol. 15, Issue 11, 5187-5196, November 2004

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The Caenorhabditis elegans Kinetochore Reorganizes at Prometaphase and in Response to Checkpoint Stimuli{boxd}

Jeffrey H. Stear * {dagger} {ddagger}, and Mark B. Roth * §

* Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA 98109; {dagger} Molecular and Cellular Biology Program, University of Washington, Seattle, WA 98195

Submitted June 14, 2004; Revised August 23, 2004; Accepted September 2, 2004
Monitoring Editor: Joseph Gall

Previous studies of the kinetochore in mammalian systems have demonstrated that this structure undergoes reorganizations after microtubule attachment or in response to activation of the spindle checkpoint. Here, we show that the Caenorhabditis elegans kinetochore displays analogous rearrangements at prometaphase, when microtubule/chromosome interactions are being established, and after exposure to checkpoint stimuli such as nocodazole or anoxia. These reorganizations are characterized by a dissociation of several kinetochore proteins, including HCP-1/CeCENP-F, HIM-10/CeNuf2, SAN-1/CeMad3, and CeBUB-1, from the centromere. We further demonstrate that at metaphase, despite having dissociated from the centromere, these reorganized kinetochore proteins maintain their associations with the metaphase plate. After checkpoint activation, these proteins are detectable as large "flares" that project out laterally from the metaphase plate. Disrupting these gene products via RNA interference results in sensitivity to checkpoint stimuli, as well as defects in the organization of chromosomes at metaphase. These phenotypes suggest that these proteins, and by extension their reorganization during mitosis, are important for mediating the checkpoint response as well as directing the assembly of the metaphase plate.


Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E04-06-0486. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E04-06-0486.

{boxd} The online version of this article contains supplementary material accessible through http://www.molbiolcell.org.

{ddagger} Present address: Max-Planck Institute for Cell Biology and Genetics, Pfotenhauerstr. 108, 01309 Dresden, Germany.

§ Corresponding author. E-mail address: mroth{at}fhcrc.org.




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