The virulence regulator VirB from Shigella flexneri uses a CTP-dependent switch mechanism to activate gene expression

Sara Jakob, Wieland Steinchen, Juri Hanßmann, Julia Rosum, Katja Langenfeld, Manuel Osorio-Valeriano, Niklas Steube, Pietro I. Giammarinaro, Georg K. A. Hochberg, Timo Glatter, Gert Bange, Andreas Diepold and Martin Thanbichler ()
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Sara Jakob: University of Marburg
Wieland Steinchen: University of Marburg
Juri Hanßmann: University of Marburg
Julia Rosum: University of Marburg
Katja Langenfeld: Max Planck Institute for Terrestrial Microbiology
Manuel Osorio-Valeriano: University of Marburg
Niklas Steube: Evolutionary Biochemistry Group, Max Planck Institute for Terrestrial Microbiology
Pietro I. Giammarinaro: University of Marburg
Georg K. A. Hochberg: University of Marburg
Timo Glatter: Max Planck Institute for Terrestrial Microbiology
Gert Bange: University of Marburg
Andreas Diepold: Max Planck Institute for Terrestrial Microbiology
Martin Thanbichler: University of Marburg

Nature Communications, 2024, vol. 15, issue 1, 1-18

Abstract: Abstract The transcriptional antisilencer VirB acts as a master regulator of virulence gene expression in the human pathogen Shigella flexneri. It binds DNA sequences (virS) upstream of VirB-dependent promoters and counteracts their silencing by the nucleoid-organizing protein H-NS. However, its precise mode of action remains unclear. Notably, VirB is not a classical transcription factor but related to ParB-type DNA-partitioning proteins, which have recently been recognized as DNA-sliding clamps using CTP binding and hydrolysis to control their DNA entry gate. Here, we show that VirB binds CTP, embraces DNA in a clamp-like fashion upon its CTP-dependent loading at virS sites and slides laterally on DNA after clamp closure. Mutations that prevent CTP-binding block VirB loading in vitro and abolish the formation of VirB nucleoprotein complexes as well as virulence gene expression in vivo. Thus, VirB represents a CTP-dependent molecular switch that uses a loading-and-sliding mechanism to control transcription during bacterial pathogenesis.

Date: 2024
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DOI: 10.1038/s41467-023-44509-z

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