Characterization of bacteriophage T4-coordinated leading- and lagging-strand synthesis on a minicircle substrate

Frank Salinas, Stephen J. Benkovic

Research output: Contribution to journalArticlepeer-review

49 Scopus citations

Abstract

The DNA replication complex of bacteriophage T4 has been assembled as a single unit on a minicircle substrate with a replication fork that permits an independent measurement of the amount of DNA synthesis on both the leading and lagging strands. The assembled replisome consists of the T4 polymerase [gene product 43 (gp43)], clamp protein (gp45), clamp loader (gp44/62), helicase (gp41), helicase accessory factor (gp59), primase (gp61), and single-stranded DNA binding protein (gp32). We demonstrate that on the minicircle the synthesis of the leading and lagging strands are coordinated and that the C-terminal domain of the gp32 protein regulates this coordination. We show that the reconstituted replisome encompasses two coupled holoenzyme complexes and present evidence that this coupling might include a gp43 homodimer interaction.

Original languageEnglish (US)
Pages (from-to)7196-7201
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume97
Issue number13
DOIs
StatePublished - Jun 20 2000

All Science Journal Classification (ASJC) codes

  • General

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