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Characterization of a Four-Component Regulatory System Controlling Bacteriocin Production in Streptococcus gallolyticus

Abstract : Bacteriocins are natural antimicrobial peptides produced by bacteria to kill closely related competitors. The opportunistic pathogen Streptococcus gallolyticus subsp. gallolyticus was recently shown to outcompete commensal enterococci of the murine microbiota under tumoral conditions thanks to the production of a two-peptide bacteriocin named gallocin. Here, we identified four genes involved in the regulatory control of gallocin in S. gallolyticus subsp. gallolyticus UCN34 that encode a histidine kinase/response regulator two-component system (BlpH/BlpR), a secreted peptide (GSP [gallocin-stimulating peptide]), and a putative regulator of unknown function (BlpS). While BlpR is a typical 243-amino-acid (aa) response regulator possessing a phospho-receiver domain and a LytTR DNA-binding domain, BlpS is a 108aa protein containing only a LytTR domain. Our results showed that the secreted peptide GSP activates the dedicated two-component system BlpH/BlpR to induce gallocin transcription. A genome-wide transcriptome analysis indicates that this regulatory system (GSP-BlpH/BlpR) is specific for bacteriocin production. Importantly, as opposed to BlpR, BlpS was shown to repress gallocin gene transcription. A conserved operator DNA sequence of 30 bp was found in all promoter regions regulated by BlpR and BlpS. Electrophoretic mobility shift assays (EMSA) and footprint assays showed direct and specific binding of BlpS and BlpR to various regulated promoter regions in a dose-dependent manner on this conserved sequence. Gallocin expression appears to be tightly controlled in S. gallolyticus subsp. gallolyticus by quorum sensing and antagonistic activity of 2 LytTR-containing proteins. Competition experiments in gut microbiota medium and 5% CO 2 to mimic intestinal conditions demonstrate that gallocin is functional under these in vivo-like conditions.
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https://hal-pasteur.archives-ouvertes.fr/pasteur-03099975
Contributor : Hugo Varet <>
Submitted on : Wednesday, January 6, 2021 - 1:52:00 PM
Last modification on : Thursday, February 25, 2021 - 2:32:21 PM
Long-term archiving on: : Wednesday, April 7, 2021 - 8:17:56 PM

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Alexis Proutière, Laurence Du Merle, Bruno Périchon, Hugo Varet, Myriam Gominet, et al.. Characterization of a Four-Component Regulatory System Controlling Bacteriocin Production in Streptococcus gallolyticus. mBio, 2021, 12 (1), ⟨10.1128/mBio.03187-20⟩. ⟨pasteur-03099975⟩

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