Scheduled conversion of replication complex architecture at replication origins of Saccharomyces cerevisiae during the cell cycle

Ryusuke Tadokoro, Masako Fujita, Hitoshi Miura, Katsuhiko Shirahige, Hiroshi Yoshikawa, Toshiki Tsurimoto, Chikashi Obuse

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Replication of DNA within Saccharomyces cerevisiae chromosomes is initiated from multiple origins, whose activation follow their own inherent time schedules during the S phase of the cell cycle. It has been demonstrated that a characteristic replicative complex (RC) that includes an origin recognition complex is formed at each origin and shifts between post- and pre-replicative states during the cell cycle. We wanted to determine whether there was an association between this shift in the state of the RC and firing events at replication origins. Time course analyses of RC architecture using LTV-footprinting with synchronously growing cells revealed that pre-replicative states at both early and late firing origins appeared simultaneously during late M phase, remained in this state during G1 phase, and converted to the post-replicative state at various times during S phase. Because the conversion of the origin footprinting profiles and origin firing, as assessed by two-dimensional gel electrophoresis, occurred concomitantly at each origin, then these two events must be closely related. However, conversion of the late firing origin occurred without actual firing. This was observed when the late origin was suppressed in clb5-deficient cells and a replication fork originating from an outside origin replicated the late origin passively. This mechanism ensures that replication at each chromosomal locus occurs only once per cell cycle by shifting existing pre-RCs to the post-RC state, when it is replicated without firing.

Original languageEnglish
Pages (from-to)15881-15889
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number18
DOIs
Publication statusPublished - May 3 2002
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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