Identification of a site required for DNA replication fork blocking activity in the rRNA gene cluster in Saccharomyces cerevisiae

Takehiko Kobayashi, Masumi Hidaka, Masafumi Nishizawa, Takashi Horiuchi

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

The yeast genome has DNA replication fork blocking sites, that we have named sog sites, in the ribosomal RNA gene (rDNA) cluster. These are located at the 3′ end of the 35S rRNA transcription unit and they block replication fork movement in a direction opposite to that of RNA polymerase I. We cloned this replication blocking site into a YEp-type plasmid and analyzed DNA replication intermediates, using two-dimensional (2D) agarose gel electrophoresis. The blocking activity remained even on a plasmid not involved in 35S rRNA transcription and inhibited fork movement in the same polar fashion as on the yeast chromosome. To define the site further, smaller fragments were subcloned into the YEp-type plasmid. A small 109 by region exhibited sog activity and was located near the enhancer region for 35S rRNA transcription. It overlaps an essential element of the recombinational hot spot HOT1.

Original languageEnglish
Pages (from-to)355-362
Number of pages8
JournalMGG Molecular & General Genetics
Volume233
Issue number3
DOIs
Publication statusPublished - 1992 Jun 1

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Keywords

  • 2 gm Plasmid
  • 2D agarose gel electrophoresis
  • Replication fork block
  • Saccharomyces cerevisiae
  • rRNA gene

ASJC Scopus subject areas

  • Genetics

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