title: Oligopeptide permease is required for expression of the Bacillus thuringiensis plcR regulon and for virulence.
abstract: PlcR is a pleiotropic regulator of virulence factors in the insect pathogen Bacillus thuringiensis and in the opportunistic human pathogen Bacillus cereus. It activates the transcription of at least 15 genes encoding extracellular proteins, including phospholipases C, proteases and enterotoxins. Expression of the plcR gene is autoregulated and activated at the onset of stationary phase. Here, we used mini-Tn10 transposition to generate a library of B. thuringiensis mutants, with the goal of characterizing genes involved in the expression of the plcR gene. Three mutant strains were identified carrying distinct mini-Tn10 insertions. The mutations impaired plcR expression and caused a deficient haemolytic phenotype, similar to the phenotype of a B. thuringiensis strain in which the plcR gene had been disrupted. The insertion sites of the three mini-Tn10 transposons mapped in a five-gene operon encoding polypeptides homologous to the components of the oligopeptide permease (Opp) system of Bacillus subtilis, and with a similar structural organization. By analogy, the five B. thuringiensis genes were designated oppA, B, C, D and F. In vitro disruption of the B. thuringiensis oppB gene reproduced the effect of the mini-Tn10 insertions (i.e. the loss of haemolytic activity) and reduced the virulence of the strain against insects. These phenotypes are similar to those of a DeltaplcR mutant. Opp is required for the import of small peptides into the cell. Therefore, plcR expression might be activated at the onset of stationary phase by the uptake of a signalling peptide acting as a quorum-sensing effector. The opp mutations impaired the sporulation efficiency of B. thuringiensis when the cells were cultured in LB medium. Thus, Opp is on the pathway that ultimately regulates Spo0A phosphorylation, as is the case in B. subtilis. However, analysis of plcR expression in DeltaoppB, Deltaspo0A and DeltaoppB Deltaspo0A mutants indicates that Opp is required for plcR expression via a Spo0A-independent mechanism.
authors.affiliation: Unité de Biochimie Microbienne, CNRS (URA2172), Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris cedex, France.
authors.forename: M
authors.initials: M
authors.lastname: Gominet
authors.forename: L
authors.initials: L
authors.lastname: Slamti
authors.forename: N
authors.initials: N
authors.lastname: Gilois
authors.forename: M
authors.initials: M
authors.lastname: Rose
authors.forename: D
authors.initials: D
authors.lastname: Lereclus
mesh_terms: D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics* / Q000472:pathogenicity*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D006460:Hemolysin Proteins / Q000235:genetics; D007814:Larva / Q000382:microbiology; D007915:Lepidoptera / Q000382:microbiology*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010740:Phospholipases / Q000235:genetics; D018085:Regulon; D017422:Sequence Analysis, DNA; D013171:Spores, Bacterial; D015534:Trans-Activators / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D010738:Type C Phospholipases / Q000235:genetics; D014774:Virulence; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D006460:Hemolysin Proteins; D026901:Membrane Transport Proteins; C099480:PlcR protein, Bacillus; C080950:Spo0A protein, Bacillus subtilis; D015534:Trans-Activators; D014157:Transcription Factors; C072861:OppB protein, Salmonella; C037230:oligopeptide permease, Bacteria; D010740:Phospholipases; D010738:Type C Phospholipases; D001616:beta-Galactosidase; D043265:Phosphatidylinositol Diacylglycerol-Lyase