Direct removal of RNA polymerase barriers to replication by accessory replicative helicases

Michelle Hawkins, Juachi U. Dimude, Jamieson A. L. Howard, Abigail Smith, Mark Dillingham, Nigel Savery, Christian J. Rudolph, Peter McGlynn

Research output: Contribution to journalArticle (Academic Journal)

4 Citations (Scopus)
169 Downloads (Pure)

Abstract

Bacterial genome duplication and transcription require simultaneous access to the same DNA template. Conflicts between the replisome and transcription machinery can lead to interruption of DNA replication and loss of genome stability. Pausing, stalling and backtracking of transcribing RNA polymerases add to this problem and present barriers to replisomes. Accessory helicases promote fork movement through nucleoprotein barriers and exist in viruses, bacteria and eukaryotes. Here we show that stalled E. coli transcription elongation complexes block reconstituted replisomes. This physiologically relevant block can be alleviated by the accessory helicase Rep or UvrD, resulting in the formation of full-length replication products. Accessory helicase action during replication-transcription collisions therefore promotes continued replication without leaving gaps in the DNA. In contrast, DinG does not promote replisome movement through stalled transcription complexes in vitro. However, our data demonstrate that DinG operates indirectly in vivo to reduce conflicts between replication and transcription. These results suggest that Rep and UvrD helicases operate on DNA at the replication fork whereas DinG helicase acts via a different mechanism.
Original languageEnglish
Article numbergkz170
Pages (from-to)5100-5113
Number of pages14
JournalNucleic Acids Research
Volume47
Issue number10
Early online date14 Mar 2019
DOIs
Publication statusPublished - 4 Jun 2019

Keywords

  • Replication
  • RNA polymerase
  • DinG
  • UvrD
  • Rep
  • Conflicts
  • Genome stability
  • Helicases
  • Transcription

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