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Published 17 March 2003. doi:10.1083/jcb.200211116
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© The Rockefeller University Press, 0021-9525/2003/3/833 $5.00
The Journal of Cell Biology, Volume 160, Number 6, 833-843

Article

 Differential kinetochore protein requirements for establishment versus propagation of centromere activity in Saccharomyces cerevisiae

Karthikeyan Mythreye and Kerry S. Bloom

Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599

Address correspondence to Kerry S. Bloom, 622 Fordham Hall, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599. Tel.: (919) 962-1182. Fax: (919) 962-8461. E-mail: kbloom{at}email.unc.edu

Dicentric chromosomes undergo a breakage–fusion–bridge cycle as a consequence of having two centromeres on the same chromatid attach to opposite spindle poles in mitosis. Suppression of dicentric chromosome breakage reflects loss of kinetochore function at the kinetochore–microtubule or the kinetochore–DNA interface. Using a conditionally functional dicentric chromosome in vivo, we demonstrate that kinetochore mutants exhibit quantitative differences in their degree of chromosome breakage. Mutations in chl4/mcm17/ctf17 segregate dicentric chromosomes through successive cell divisions without breakage, indicating that only one of the two centromeres is functional. Centromere DNA introduced into the cell is unable to promote kinetochore assembly in the absence of CHL4. In contrast, established centromeres retain their segregation capacity for greater than 25 generations after depletion of Chl4p. The persistent mitotic stability of established centromeres reveals the presence of an epigenetic component in kinetochore segregation. Furthermore, this study identifies Chl4p in the initiation and specification of a heritable chromatin state.

Key Words: yeast; centromere; kinetochore; chromatin; epigenetic

* Abbreviation used in this paper: ChIP, chromatin immunoprecipitation.


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