title: The multicopy sRNA LhrC controls expression of the oligopeptide-binding protein OppA in Listeria monocytogenes.
abstract: Listeria monocytogenes is the causative agent of the foodborne disease listeriosis. During infection, L. monocytogenes produces an array of non-coding RNAs, including the multicopy sRNA LhrC. These five, nearly identical sRNAs are highly induced in response to cell envelope stress and target the virulence adhesin lapB at the post-transcriptional level. Here, we demonstrate that LhrC controls expression of additional genes encoding cell envelope-associated proteins with virulence function. Using transcriptomics and proteomics, we identified a set of genes affected by LhrC in response to cell envelope stress. Three targets were significantly down-regulated by LhrC at both the RNA and protein level: lmo2349, tcsA and oppA. All three genes encode membrane-associated proteins: A putative substrate binding protein of an amino acid ABC transporter (Lmo2349); the CD4+ T cell-stimulating antigen TcsA, and the oligopeptide binding protein OppA, of which the latter 2 are required for full virulence of L. monocytogenes. For OppA, we show that LhrC acts by direct base paring to the ribosome binding site of the oppA mRNA, leading to an impediment of its translation and a decreased mRNA level. The sRNA-mRNA interaction depends on 2 of 3 CU-rich regions in LhrC allowing binding of 2 oppA mRNAs to a single LhrC molecule. Finally, we found that LhrC contributes to infection in macrophage-like cells. These findings demonstrate a central role for LhrC in controlling the level of OppA and other virulence-associated cell envelope proteins in response to cell envelope stress.
authors.affiliation: a Department of Biochemistry and Molecular Biology ; University of Southern Denmark ; Odense , Denmark.|b Institute for Microbiology; Ernst-Moritz-Arndt-University Greifswald ; Greifswald , Germany.|d These authors equally contributed to this work.
authors.forename: Susanne
authors.initials: S
authors.lastname: Sievers
authors.affiliation: a Department of Biochemistry and Molecular Biology ; University of Southern Denmark ; Odense , Denmark.|d These authors equally contributed to this work.
authors.forename: Anja
authors.initials: A
authors.lastname: Lund
authors.affiliation: a Department of Biochemistry and Molecular Biology ; University of Southern Denmark ; Odense , Denmark.
authors.forename: Pilar
authors.initials: P
authors.lastname: Menendez-Gil
authors.affiliation: a Department of Biochemistry and Molecular Biology ; University of Southern Denmark ; Odense , Denmark.
authors.forename: Aaraby
authors.initials: A
authors.lastname: Nielsen
authors.affiliation: a Department of Biochemistry and Molecular Biology ; University of Southern Denmark ; Odense , Denmark.
authors.forename: Maria
authors.initials: M
authors.lastname: Storm Mollerup
authors.affiliation: a Department of Biochemistry and Molecular Biology ; University of Southern Denmark ; Odense , Denmark.
authors.forename: Stine
authors.initials: S
authors.lastname: Lambert Nielsen
authors.affiliation: a Department of Biochemistry and Molecular Biology ; University of Southern Denmark ; Odense , Denmark.
authors.forename: Pernille
authors.initials: P
authors.lastname: Buch Larsson
authors.affiliation: a Department of Biochemistry and Molecular Biology ; University of Southern Denmark ; Odense , Denmark.
authors.forename: Jonas
authors.initials: J
authors.lastname: Borch-Jensen
authors.affiliation: c Department of Molecular Biology ; Umeå University ; Umeå , Sweden.
authors.forename: Jörgen
authors.initials: J
authors.lastname: Johansson
authors.affiliation: a Department of Biochemistry and Molecular Biology ; University of Southern Denmark ; Odense , Denmark.
authors.forename: Birgitte Haahr
authors.initials: BH
authors.lastname: Kallipolitis
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D018628:Gene Dosage; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D008074:Lipoproteins / Q000235:genetics* / Q000378:metabolism; D008089:Listeria monocytogenes / Q000235:genetics* / Q000378:metabolism; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D009154:Mutation; D009690:Nucleic Acid Conformation; D059372:Nucleotide Motifs; D009876:Operon; D020543:Proteome; D040901:Proteomics / Q000379:methods; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D058727:RNA, Small Untranslated / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013312:Stress, Physiological / Q000235:genetics; D059467:Transcriptome
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D008074:Lipoproteins; D020543:Proteome; D012333:RNA, Messenger; D058727:RNA, Small Untranslated; C046506:oligopeptide-binding protein, bacteria
keywords: Listeria monocytogenes; cell envelope stress; proteomics; sRNA; transcriptomics