A DNA replication-arrest site RTS1 regulates imprinting by determining the direction of replication at mat1 in S. pombe

  • Dalgaard J
  • Klar A
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Abstract

Mating-type switching in Schizosaccharomyces pombe involves a strand-specific, alkali-labile imprint at the mat1 (mating-type) locus. The imprint is synthesized during replication in a swi1, swi3, and polyinerase alpha (swi7) dependent manner and is dependent on mat1 being replicated in a specific direction. Here we show that the direction of replication at mat1 is controlled by a cis-acting polar terminator of replication (RTS1). Two-dimensional gel analysis of replication intermediates reveals that RTS1 only terminates replication forks moving in the centromere-distal direction. A genetic analysis shows that RTS1 optimizes the imprinting process. Transposing the RTS1 element to the distal side of mat1 abolishes imprinting of the native mat1 allele but restores imprinting of an otherwise unimprinted inverted mat1 allele. These data provide conclusive evidence for the ''direction of replication model'' that explains the asymmetrical switching pattern of S. Pombe, and identify a DNA replication-arrest element implicated in a developmental process. Such elements could play a more general role during development and differentiation in higher eukaryotes by regulating the direction of DNA replication at key loci.

Author-supplied keywords

  • Differentiation
  • Imprinting
  • Mating-type switching
  • Schizosaccharomyces pombe
  • Termination of DNA replication

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Authors

  • Jacob Z. Dalgaard

  • A. J S Klar

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