DksA and DNA Double-Strand Break Repair

Curr Genet. 2019 Dec;65(6):1297-1300. doi: 10.1007/s00294-019-00983-x. Epub 2019 May 10.

Abstract

We use genetic assays to suggest that transcription-coupled repair or new origin formation in Escherichia coli involves removal of RNAP to create an RNA primer for DNA synthesis. Transcription factor DksA was shown to play a role in numerous reactions involving RNA polymerase. Some, but not all, of the activities of DksA at promoters or during transcription elongation require (p)ppGpp. In addition to its role during transcription, DksA is also involved in maintaining genome integrity. Cells lacking DksA are sensitive to multiple DNA damaging agents including UV light, ionizing radiation, mitomycin C, and nalidixic acid. Here, we focus on two recent studies addressing the importance of DksA in the repair of double-strand breaks (DSBs), one by Sivaramakrishnan et al. (Nature 550:214-218, 2017) and one originating in our laboratory, Myka et al. (Mol Microbiol 111:1382-1397. https://doi.org/10.1111/mmi.14227 , 2019). It appears that depending on the type and possibly location of DNA damage, DksA can play either a passive or an active role in DSB repair. The passive role relies on exclusion of anti-backtracking factors from the RNAP secondary channel. The exact mechanism of active DksA-mediated DNA repair is unknown. However, DksA was proposed to destabilize transcription complexes, thus clearing the way for recombination and DNA repair. Based on the requirement for DksA, both in repair of DSBs and the R-loop-dependent formation of new origins of DNA replication, we propose that DksA may allow for removal of RNAP without unwinding of the RNA:DNA hybrid, which can then be extended by a DNA polymerase. This mechanism obviates the need for RNAP backtracking to repair damaged DNA.

Keywords: (p)ppGpp; Anti-backtracking factors; Double-strand break repair; Nalidixic acid; Phleomycin; Replication–transcription conflicts; Transcription factor DksA; Type II topoisomerase.

Publication types

  • Review

MeSH terms

  • DNA Breaks, Double-Stranded* / drug effects
  • DNA Repair*
  • DNA Topoisomerases, Type II / metabolism
  • DNA-Directed RNA Polymerases / metabolism
  • Escherichia coli / genetics*
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Guanosine Pentaphosphate / metabolism
  • Nalidixic Acid / pharmacology
  • Phleomycins / pharmacology
  • Promoter Regions, Genetic
  • Transcription Factors / genetics

Substances

  • Escherichia coli Proteins
  • Phleomycins
  • Transcription Factors
  • dksA protein, E coli
  • Guanosine Pentaphosphate
  • Nalidixic Acid
  • DNA-Directed RNA Polymerases
  • DNA Topoisomerases, Type II