Thermus thermophilus Argonaute Functions in the Completion of DNA Replication

Cell. 2020 Sep 17;182(6):1545-1559.e18. doi: 10.1016/j.cell.2020.07.036. Epub 2020 Aug 25.

Abstract

In many eukaryotes, Argonaute proteins, guided by short RNA sequences, defend cells against transposons and viruses. In the eubacterium Thermus thermophilus, the DNA-guided Argonaute TtAgo defends against transformation by DNA plasmids. Here, we report that TtAgo also participates in DNA replication. In vivo, TtAgo binds 15- to 18-nt DNA guides derived from the chromosomal region where replication terminates and associates with proteins known to act in DNA replication. When gyrase, the sole T. thermophilus type II topoisomerase, is inhibited, TtAgo allows the bacterium to finish replicating its circular genome. In contrast, loss of gyrase and TtAgo activity slows growth and produces long sausage-like filaments in which the individual bacteria are linked by DNA. Finally, wild-type T. thermophilus outcompetes an otherwise isogenic strain lacking TtAgo. We propose that the primary role of TtAgo is to help T. thermophilus disentangle the catenated circular chromosomes generated by DNA replication.

Keywords: Argonaute; DNA replication; RNA silencing; Thermus thermophilus; TtAgo; decatenation; gyrase; pAGO; terminus of replication; topoisomerase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Argonaute Proteins / genetics
  • Argonaute Proteins / metabolism*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Cell Survival / drug effects
  • Cell Survival / genetics
  • Chromosomes / metabolism
  • Ciprofloxacin / pharmacology
  • DNA / genetics
  • DNA / metabolism*
  • DNA Gyrase / metabolism*
  • DNA Replication / drug effects
  • DNA Replication / genetics*
  • Endonucleases / metabolism
  • Microscopy, Electron, Scanning
  • Microscopy, Electron, Transmission
  • Models, Molecular
  • Recombinant Proteins
  • Recombination, Genetic / drug effects
  • Recombination, Genetic / genetics
  • Single Molecule Imaging
  • Tandem Mass Spectrometry
  • Thermus thermophilus / genetics
  • Thermus thermophilus / growth & development
  • Thermus thermophilus / metabolism*
  • Thermus thermophilus / ultrastructure
  • Topoisomerase II Inhibitors / pharmacology

Substances

  • Argonaute Proteins
  • Bacterial Proteins
  • Recombinant Proteins
  • Topoisomerase II Inhibitors
  • Ciprofloxacin
  • DNA
  • Endonucleases
  • DNA Gyrase