RNA‐binding protein Hfq downregulates locus of enterocyte effacement‐encoded regulators independent of small regulatory RNA in enterohemorrhagic Escherichia coli

• Published in: Molecular microbiology Vol. 117; no. 1; pp. 86 - 101
• Main Authors: Sudo, Naoki, Lee, Kenichi, Sekine, Yasuhiko, Ohnishi, Makoto, Iyoda, Sunao
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.01.2022
• Subjects: E coli; EHEC; Enterohemorrhagic Escherichia coli - genetics; Enterohemorrhagic Escherichia coli - growth & development; Enterohemorrhagic Escherichia coli - pathogenicity; Escherichia coli; Escherichia coli Infections - microbiology; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Gene expression; Gene Expression Regulation, Bacterial - genetics; Genetic suppression; Hfq; Host Factor 1 Protein - genetics; Host Factor 1 Protein - metabolism; Humans; Loci; locus of enterocyte effacement; Mutation; pathogenicity; Phosphoproteins - genetics; Phosphoproteins - metabolism; Post-transcription; posttranscriptional regulation; Protein Biosynthesis; Proteins; Ribonucleic acid; RNA; RNA, Bacterial - genetics; RNA, Messenger - genetics; RNA, Small Untranslated - genetics; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; Trans-Activators - genetics; Trans-Activators - metabolism; Translation; Translation initiation; Type III Secretion Systems; type three secretion system; Virulence; posttranscriptional regulation; EHEC; pathogenicity; Hfq; type three secretion system; locus of enterocyte effacement
• ISSN: 0950-382X; 1365-2958; 1365-2958
• DOI: 10.1111/mmi.14799

Enterohemorrhagic Escherichia coli (EHEC) causes severe human diseases worldwide. The type 3 secretion system and effector proteins are essential for EHEC infection, and are encoded by the locus of enterocyte effacement (LEE). RNA‐binding protein Hfq is essential for small regulatory RNA (sRNA)‐mediated regulation at a posttranscriptional level and full virulence of many pathogenic bacteria. Although two early studies indicated that Hfq represses LEE expression by posttranscriptionally controlling the expression of genes grlRA and/or ler, both of which encode LEE regulators mediating a positive regulatory loop, the detailed molecular mechanism and biological significance remain unclear. Herein, we show that LEE overexpression was caused by defective RNA‐binding activity of the Hfq distal face, which posttranscriptionally represses grlA and ler expression. In vitro analyses revealed that the Hfq distal face directly binds near the translational initiation site of grlA and ler mRNAs, and inhibits their translation. Taken together, we conclude that Hfq inhibits grlA and ler translation by binding their mRNAs through the distal face in an sRNA‐independent manner. Additionally, we show that Hfq‐mediated repression of LEE is critical for normal EHEC growth because all suppressor mutations that restored the growth defect in the hfq mutant abolished hfq deletion‐induced overexpression of LEE. The locus of enterocyte effacement (LEE) is an important virulence gene region in enterohemorrhagic Escherichia coli. Although LEE expression is regulated by RNA‐binding protein Hfq, which is essential for small regulatory RNA (sRNA)‐mediated regulation, the mechanism is uncertain. Herein, we revealed that the distal RNA‐binding surface of Hfq posttranscriptionally inhibits expression of LEE regulators through an sRNA‐independent mechanism, resulting in strong repression of LEE expression.