Protein-DNA complexes are the primary sources of replication fork pausing in Escherichia coli

Milind K. Gupta, Colin P. Guy, Joseph T. P. Yeeles, John Atkinson, Hazel Bell, Robert G. Lloyd, Kenneth J. Marians, Peter McGlynn*

*Corresponding author for this work

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Replication fork pausing drives genome instability, because any loss of paused replisome activity creates a requirement for reloading of the replication machinery, a potentially mutagenic process. Despite this importance, the relative contributions to fork pausing of different replicative barriers remain unknown. We show here that Deinococcus radiodurans RecD2 helicase inactivates Escherichia coli replisomes that are paused but still functional in vitro, preventing continued fork movement upon barrier removal or bypass, but does not inactivate elongating forks. Using RecD2 to probe replisome pausing in vivo, we demonstrate that most pausing events do not lead to replisome inactivation, that transcription complexes are the primary sources of this pausing, and that an accessory replicative helicase is critical for minimizing the frequency and/or duration of replisome pauses. These findings reveal the hidden potential for replisome inactivation, and hence genome instability, inside cells. They also demonstrate that efficient chromosome duplication requires mechanisms that aid resumption of replication by paused replisomes, especially those halted by protein-DNA barriers such as transcription complexes.

Original languageEnglish
Pages (from-to)7252-7257
Number of pages6
JournalPNAS
Volume110
Issue number18
DOIs
Publication statusPublished - 30 Apr 2013

Keywords

  • DNA repair
  • genome stability
  • rep
  • RNA polymerase
  • recombination
  • units in-vivo
  • RNA-polymerase
  • lagging-strand
  • direct restart
  • transcription
  • replisome
  • helicase
  • progression
  • chromosome

Cite this

Gupta, M. K., Guy, C. P., Yeeles, J. T. P., Atkinson, J., Bell, H., Lloyd, R. G., ... McGlynn, P. (2013). Protein-DNA complexes are the primary sources of replication fork pausing in Escherichia coli. PNAS, 110(18), 7252-7257. https://doi.org/10.1073/pnas.1303890110

Protein-DNA complexes are the primary sources of replication fork pausing in Escherichia coli. / Gupta, Milind K.; Guy, Colin P.; Yeeles, Joseph T. P.; Atkinson, John; Bell, Hazel; Lloyd, Robert G.; Marians, Kenneth J.; McGlynn, Peter.

In: PNAS, Vol. 110, No. 18, 30.04.2013, p. 7252-7257.

Research output: Contribution to journalArticle

Gupta, MK, Guy, CP, Yeeles, JTP, Atkinson, J, Bell, H, Lloyd, RG, Marians, KJ & McGlynn, P 2013, 'Protein-DNA complexes are the primary sources of replication fork pausing in Escherichia coli', PNAS, vol. 110, no. 18, pp. 7252-7257. https://doi.org/10.1073/pnas.1303890110
Gupta MK, Guy CP, Yeeles JTP, Atkinson J, Bell H, Lloyd RG et al. Protein-DNA complexes are the primary sources of replication fork pausing in Escherichia coli. PNAS. 2013 Apr 30;110(18):7252-7257. https://doi.org/10.1073/pnas.1303890110
Gupta, Milind K. ; Guy, Colin P. ; Yeeles, Joseph T. P. ; Atkinson, John ; Bell, Hazel ; Lloyd, Robert G. ; Marians, Kenneth J. ; McGlynn, Peter. / Protein-DNA complexes are the primary sources of replication fork pausing in Escherichia coli. In: PNAS. 2013 ; Vol. 110, No. 18. pp. 7252-7257.
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