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Chromosome Research

, Volume 11, Issue 2, pp 137–146 | Cite as

RNA interference is required for normal centromere function infission yeast

  • Tom Volpe
  • Vera Schramke
  • Georgina L. Hamilton
  • Sharon A. White
  • Grace Teng
  • Robert A. Martienssen
  • Robin C. Allshire
Article

Abstract

In plants, animals and fungi, active centromeres are associated with arrays of repetitive DNA sequences. The outer repeats at fission yeast (Schizosaccharomyces pombe) centromeres are heterochromatic and are required for the assembly of an active centromere. Components of the RNA interference (RNAi) machinery process transcripts derived from these repeats and mediate the formation of silent chromatin. A subfragment of the repeat (dg) is known to induce silencing of marker genes at euchromatic sites and is required for centromere formation. We show that the RNAi components, Argonaute (Ago1), Dicer (Dcr1) and RNA-dependent RNA polymerase (Rdp1), are required to maintain silencing, lysine 9 methylation of histone H3 and association of Swi6 via this dg ectopic silencer. Deletion of Ago1, Dcr1 or Rdp1 disrupts chromosome segregation leading to a high incidence of lagging chromosomes on late anaphase spindles and sensitivity to a microtubule poison. Analysis of dg transcription indicates that csp mutants, previously shown to abrogate centromere silencing and chromosome segregation, are also defective in the regulation of non-coding centromeric RNAs. In addition, histone H3 lysine 9 methylation at, and recruitment of Swi6 and cohesin to, centromeric repeats is disrupted in these mutants. Thus the formation of silent chromatin on dg repeats and the development of a fully functional centromere is dependent on RNAi.

centromere chromosome segregation kinetochore RNAi Schizosaccharomyces pombe 

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References

  1. Allshire RC (1996) Transcriptional silencing in fission yeast: A manifestation of higher order chromosome structure and functions. In: Russo E, Martienssen R, eds. Epigenetic Mechanisms of Gene Regulation. New York, USA: CSHL Press, pp 443-466.Google Scholar
  2. Allshire RC, Nimmo ER, Ekwall K, Javerzat J-P, Cranston G (1995) Mutations derepressing silent centromeric domains in fission yeast disrupt chromosome segregation. Genes Dev 9: 218-233.Google Scholar
  3. Ayoub N, Goldshmidt I, Lyakhovetsky R, Cohen A (2000) A fission yeast repression element cooperates with centromere-like sequences and defines a mat silent domain boundary. Genetics 156: 983-994.Google Scholar
  4. Bannister AJ, Zegerman P, Partridge JF et al. (2001) Selective recognition of methylated lysine 9 on histone H3 by the HP1 chromo domain. Nature 410: 120-124.Google Scholar
  5. Bernard P, Allshire RC (2002) Centromeres become unstuck without heterochromatin. Trends Cell Biol 12: 419-424.Google Scholar
  6. Bernard P, Maure JF, Partridge JF, Genier S, Javerzat J-P, Allshire RC (2001) Requirement of heterochromatin for cohesion at centromeres. Science 294: 2539-2542.Google Scholar
  7. Ekwall K, Javerzat JP, Lorentz A, Schmidt H, Cranston G, Allshire RC (1995) The chromodomain protein Swi6: a key component at fission yeast centromeres. Science 269: 1429-1431.Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Tom Volpe
    • 1
  • Vera Schramke
    • 2
  • Georgina L. Hamilton
    • 2
  • Sharon A. White
    • 2
  • Grace Teng
    • 1
  • Robert A. Martienssen
    • 1
  • Robin C. Allshire
    • 2
  1. 1.Cold Spring Harbor LaboratoryNew YorkUSA
  2. 2.The Wellcome Trust Centre for Cell BiologyUniversity of Edinburgh, Kings′ BuildingsEdinburghUK

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