Coordinated leading and lagging strand DNA synthesis on a minicircular template

Joonsoo Lee, Paul D. Chastain, Takahiro Kusakabe, Jack D. Griffith, Charles C. Richardson

Research output: Contribution to journalArticlepeer-review

114 Citations (Scopus)

Abstract

The coordinated synthesis of both leading and lagging DNA strands is thought to involve a dimeric DNA polymerase and a looping of the lagging strand so that both strands can be synthesized in the same direction. We have constructed a minicircle with a replication fork that permits an assessment of the stoichiometry of the proteins and a measurement of the synthesis of each strand. The replisome consisting of bacteriophage T7 DNA polymerase, helicase, primase, and single-stranded DNA-binding protein mediates coordinated replication. The criteria for coordination are fulfilled : (1) a replication loop is formed, (2) leading and lagging strand synthesis are coupled, (3) the lagging strand polymerase recycles from one Okazaki fragment to another, and (4) the length of Okazaki fragments is regulated. T7 single-stranded DNA-binding protein is essential for coordination.

Original languageEnglish
Pages (from-to)1001-1010
Number of pages10
JournalMolecular Cell
Volume1
Issue number7
DOIs
Publication statusPublished - Jun 1998

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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