Rescue of stalled replication forks by RecG: simultaneous translocation on the leading and lagging strand templates supports an active DNA unwinding model of fork reversal and Holliday junction formation

Peter McGlynn, R. G. Lloyd

Research output: Contribution to journalArticle

146 Citations (Scopus)

Abstract

Modification of damaged replication forks is emerging as a crucial factor for efficient chromosomal duplication and the avoidance of genetic instability. The RecG helicase of Escherichia coil, which is involved in recombination and DNA repair, has been postulated to act on stalled replication forks to promote replication restart via the formation of a four-stranded (Holliday) junction. Here we show that RecC can actively unwind the leading and tagging strand arms of model replication fork structures in vitro. Unwinding is achieved in each case by simultaneous interaction with and translocation along both the leading and lagging strand templates at a fork. Disruption of either of these interactions dramatically inhibits unwinding of the opposing duplex arm. Thus, RecG translocates simultaneously along two DNA strands, one with 5 ' -3 ' and the otherwith 3 ' -5 ' polarity. The unwinding of both nascent strands at a damaged fork, and their subsequent annealing to form a Holliday junction, may explain the ability of RecC to promote replication restart. Moreover, the preferential binding of partial forks lacking a leading strand suggests that RecC may have the ability to target stalled replication intermediates in vivo in which lagging strand synthesis has continued beyond the leading strand.

Original languageEnglish
Pages (from-to)8227-8234
Number of pages7
JournalPNAS
Volume98
Issue number15
DOIs
Publication statusPublished - 2001

Keywords

  • DNA repair
  • recombination
  • helicases
  • BRANCH MIGRATION PROTEIN
  • ESCHERICHIA-COLI
  • HELICASE ACTIVITY
  • PYRIMIDINE DIMER
  • MECHANISM
  • PRIA
  • RECOMBINATION
  • REPAIR
  • TRANSCRIPTION
  • MODULATION

Cite this

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title = "Rescue of stalled replication forks by RecG: simultaneous translocation on the leading and lagging strand templates supports an active DNA unwinding model of fork reversal and Holliday junction formation",
abstract = "Modification of damaged replication forks is emerging as a crucial factor for efficient chromosomal duplication and the avoidance of genetic instability. The RecG helicase of Escherichia coil, which is involved in recombination and DNA repair, has been postulated to act on stalled replication forks to promote replication restart via the formation of a four-stranded (Holliday) junction. Here we show that RecC can actively unwind the leading and tagging strand arms of model replication fork structures in vitro. Unwinding is achieved in each case by simultaneous interaction with and translocation along both the leading and lagging strand templates at a fork. Disruption of either of these interactions dramatically inhibits unwinding of the opposing duplex arm. Thus, RecG translocates simultaneously along two DNA strands, one with 5 ' -3 ' and the otherwith 3 ' -5 ' polarity. The unwinding of both nascent strands at a damaged fork, and their subsequent annealing to form a Holliday junction, may explain the ability of RecC to promote replication restart. Moreover, the preferential binding of partial forks lacking a leading strand suggests that RecC may have the ability to target stalled replication intermediates in vivo in which lagging strand synthesis has continued beyond the leading strand.",
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author = "Peter McGlynn and Lloyd, {R. G.}",
year = "2001",
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TY - JOUR

T1 - Rescue of stalled replication forks by RecG: simultaneous translocation on the leading and lagging strand templates supports an active DNA unwinding model of fork reversal and Holliday junction formation

AU - McGlynn, Peter

AU - Lloyd, R. G.

PY - 2001

Y1 - 2001

N2 - Modification of damaged replication forks is emerging as a crucial factor for efficient chromosomal duplication and the avoidance of genetic instability. The RecG helicase of Escherichia coil, which is involved in recombination and DNA repair, has been postulated to act on stalled replication forks to promote replication restart via the formation of a four-stranded (Holliday) junction. Here we show that RecC can actively unwind the leading and tagging strand arms of model replication fork structures in vitro. Unwinding is achieved in each case by simultaneous interaction with and translocation along both the leading and lagging strand templates at a fork. Disruption of either of these interactions dramatically inhibits unwinding of the opposing duplex arm. Thus, RecG translocates simultaneously along two DNA strands, one with 5 ' -3 ' and the otherwith 3 ' -5 ' polarity. The unwinding of both nascent strands at a damaged fork, and their subsequent annealing to form a Holliday junction, may explain the ability of RecC to promote replication restart. Moreover, the preferential binding of partial forks lacking a leading strand suggests that RecC may have the ability to target stalled replication intermediates in vivo in which lagging strand synthesis has continued beyond the leading strand.

AB - Modification of damaged replication forks is emerging as a crucial factor for efficient chromosomal duplication and the avoidance of genetic instability. The RecG helicase of Escherichia coil, which is involved in recombination and DNA repair, has been postulated to act on stalled replication forks to promote replication restart via the formation of a four-stranded (Holliday) junction. Here we show that RecC can actively unwind the leading and tagging strand arms of model replication fork structures in vitro. Unwinding is achieved in each case by simultaneous interaction with and translocation along both the leading and lagging strand templates at a fork. Disruption of either of these interactions dramatically inhibits unwinding of the opposing duplex arm. Thus, RecG translocates simultaneously along two DNA strands, one with 5 ' -3 ' and the otherwith 3 ' -5 ' polarity. The unwinding of both nascent strands at a damaged fork, and their subsequent annealing to form a Holliday junction, may explain the ability of RecC to promote replication restart. Moreover, the preferential binding of partial forks lacking a leading strand suggests that RecC may have the ability to target stalled replication intermediates in vivo in which lagging strand synthesis has continued beyond the leading strand.

KW - DNA repair

KW - recombination

KW - helicases

KW - BRANCH MIGRATION PROTEIN

KW - ESCHERICHIA-COLI

KW - HELICASE ACTIVITY

KW - PYRIMIDINE DIMER

KW - MECHANISM

KW - PRIA

KW - RECOMBINATION

KW - REPAIR

KW - TRANSCRIPTION

KW - MODULATION

U2 - 10.1073/pnas.111008698

DO - 10.1073/pnas.111008698

M3 - Article

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SP - 8227

EP - 8234

JO - PNAS

JF - PNAS

SN - 0027-8424

IS - 15

ER -