Chromodomain Protein Swi6-mediated Role of DNA Polymerase α in Establishment of Silencing in Fission Yeast

Shakil Ahmed, Sharanjot Saini, Sumit Arora, Jagmohan Singh

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

Although DNA replication has been thought to play an important role in the silencing of mating type loci in Saccharomyces cerevisiae, recent studies indicate that silencing can be decoupled from replication. In Schizosaccharomyces pombe, mating type silencing is brought about by the trans-acting proteins, namely Swi6, Clr1-Clr4, and Rhp6, in cooperation with the cis-acting silencers. The latter contain an autonomous replication sequence, suggesting that DNA replication may be critical for silencing in S. pombe. To investigate the connection between DNA replication and silencing in S. pombe, we analyzed several temperature-sensitive mutants of DNA polymerase α. We find that one such mutant, swi7H4, exhibits silencing defects at mat, centromere, and telomere loci. This effect is independent of the checkpoint and replication defects of the mutant. Interestingly, the extent of the silencing defect in the swi7H4 mutant at the silent mat2 locus is further enhanced in absence of the cis-acting, centromere-proximal silencer. The chromodomain protein Swi6, which is required for silencing and is localized to mat and other heterochromatin loci, interacts with DNA polymerase α in vivo and in vitro in wild type cells. However, it does not interact with the mutant polα and is delocalized away from the silent mat loci in the mutant. Our results demonstrate a role of DNA polymerase α in the establishment of silencing. We propose a recruitment model for the coupling of DNA replication with the establishment of silencing by the chromodomain protein Swi6, which may be applicable to higher eukaryotes.

Original languageEnglish (US)
Pages (from-to)47814-47821
Number of pages8
JournalJournal of Biological Chemistry
Volume276
Issue number51
DOIs
StatePublished - Dec 21 2001

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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