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MBC in Press, published online ahead of print September 15, 2004
Mol. Biol. Cell 10.1091/mbc.E04-06-0486

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Submitted on June 14, 2004
Revised on August 23, 2004
Accepted on September 2, 2004

The C. elegans Kinetochore Reorganizes at Prometaphase and in Response to Checkpoint Stimuli

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

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

Monitoring Editor: Joseph Gall

Previous studies of the kinetochore in mammalian systems have demonstrated that this structure undergoes reorganizations following microtubule attachment or in response to activation of the spindle checkpoint. Here we show that the C. elegans kinetochore displays analogous rearrangements at prometaphase, when microtubule/chromosome interactions are being established, and following 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. Following checkpoint activation, these proteins are detectable as large "flares" that project out laterally from the metaphase plate. Disrupting these gene products via RNAi 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.

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

{sect}Corresponding author. E-mail: mroth{at}fred.fhcrc.org




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