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

Frank Salinas, Stephen Benkovic

Research output: Contribution to journalArticle

49 Citations (Scopus)

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

Fingerprint

Bacteriophage T4
DNA Primase
Holoenzymes
DNA-Binding Proteins
DNA Replication
Genes
DNA
Proteins
Protein Domains

All Science Journal Classification (ASJC) codes

  • General

Cite this

@article{5fd44dffcf2049438ad8637b73026f26,
title = "Characterization of bacteriophage T4-coordinated leading- and lagging-strand synthesis on a minicircle substrate",
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.",
author = "Frank Salinas and Stephen Benkovic",
year = "2000",
month = "6",
day = "20",
doi = "10.1073/pnas.97.13.7196",
language = "English (US)",
volume = "97",
pages = "7196--7201",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "13",

}

Characterization of bacteriophage T4-coordinated leading- and lagging-strand synthesis on a minicircle substrate. / Salinas, Frank; Benkovic, Stephen.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 97, No. 13, 20.06.2000, p. 7196-7201.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Characterization of bacteriophage T4-coordinated leading- and lagging-strand synthesis on a minicircle substrate

AU - Salinas, Frank

AU - Benkovic, Stephen

PY - 2000/6/20

Y1 - 2000/6/20

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0034691148&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034691148&partnerID=8YFLogxK

U2 - 10.1073/pnas.97.13.7196

DO - 10.1073/pnas.97.13.7196

M3 - Article

VL - 97

SP - 7196

EP - 7201

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 13

ER -