Structural basis for the inhibition of RecBCD by Gam and its synergistic antibacterial effect with quinolones

Martin Wilkinson, Lucy Troman, Wan Nur Ismah Wan Ahmad Kamil, Yuriy Chaban, Matthew B Avison, Mark Simon Dillingham*, Dale B Wigley

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)

10 Citations (Scopus)
319 Downloads (Pure)

Abstract

Our previous paper (Wilkinson et al, 2016) used high-resolution cryo-electron microscopy to solve the structure of the Escherichia coli RecBCD complex, which acts in both the repair of double-stranded DNA breaks and the degradation of bacteriophage DNA. To counteract the latter activity, bacteriophage λ encodes a small protein inhibitor called Gam that binds to RecBCD and inactivates the complex. Here, we show that Gam inhibits RecBCD by competing at the DNA-binding site. The interaction surface is extensive and involves molecular mimicry of the DNA substrate. We also show that expression of Gam in E. coli or Klebsiella pneumoniae increases sensitivity to fluoroquinolones; antibacterials that kill cells by inhibiting topoisomerases and inducing double-stranded DNA breaks. Furthermore, fluoroquinolone-resistance in K. pneumoniae clinical isolates is reversed by expression of Gam. Together, our data explain the synthetic lethality observed between topoisomerase-induced DNA breaks and the RecBCD gene products, suggesting a new co-antibacterial strategy.

Original languageEnglish
Article numbere22963
Number of pages12
JournaleLife
Volume5
DOIs
Publication statusPublished - 23 Dec 2016

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