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The Nucleoid Proteins Fis and IHF Positively Regulate the Gene Expression of Operons Responsible for Producing the Cytotoxins Tilimycin and Tilivalline in Klebsiella oxytoca
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The Nucleoid Proteins Fis and IHF Positively Regulate the Gene Expression of Operons Responsible for Producing the Cytotoxins Tilimycin and Tilivalline in Klebsiella oxytoca
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Journal Article
The Nucleoid Proteins Fis and IHF Positively Regulate the Gene Expression of Operons Responsible for Producing the Cytotoxins Tilimycin and Tilivalline in Klebsiella oxytoca
2025
Overview
causes antibiotic-associated hemorrhagic colitis due to the production of the enterotoxins tilimycin and tilivalline. These toxins are synthesized by enzymes encoded in the
and NRPS operons, which are expressed divergently. This study investigated how the nucleoid-associated proteins Fis and IHF regulate these operons and influence the production of enterotoxins.
We used reverse transcription quantitative PCR (RT-qPCR) to assess the role of Fis and IHF in the transcription of the
and NRPS operons. Electrophoretic mobility shift assays (EMSAs) were used to examine the binding of Fis and IHF to the regulatory region. Additionally, Caco-2 viability assays were performed using cells infected with WT, mutant, and complemented strains.
RT-qPCR demonstrated that deletions of
or
/
significantly reduced operon expression. EMSA confirmed that Fis and IHF bind specifically to the regulatory region between the
and NRPS operons. Viability assays in Caco-2 epithelial cells indicated increased host cell survival when exposed to the deletion mutants. Genetic complementation restored both transcription levels and cytotoxicity.
Fis and IHF are positive regulators of the
and NRPS operons, enhancing the production of tilimycin and tilivalline. These findings highlight the potential of targeting Fis and IHF for therapeutic intervention in antibiotic-associated hemorrhagic colitis.
Publisher
John Wiley & Sons, Inc,Wiley
Subject
