title: Regulation of sporulation initiation by NprR and its signaling peptide NprRB: molecular recognition and conformational changes.
abstract: NprR belongs to the RNPP family of quorum-sensing receptors, a group of intracellular regulators activated directly by signaling oligopeptides in Gram-positive bacteria. In Bacillus thuringiensis (Bt), nprR is located in a transcriptional cassette with nprRB that codes for the precursor of the signaling peptide NprRB. NprR is a transcriptional regulator activated by binding of reimported NprRB; however, several reports suggest that NprR also participates in sporulation but the mechanism is unknown. Our in silico results, based on the structural similarity between NprR from Bt and Spo0F-binding Rap proteins from Bacillus subtilis, suggested that NprR could bind Spo0F to modulate the sporulation phosphorelay in Bt. Deletion of nprR-nprRB cassette from Bt caused a delay in sporulation and defective trigger of the Spo0A∼P-activated genes spoIIA and spoIIIG. The DNA-binding domain of NprR was not necessary for this second function, since truncated NprRΔHTH together with nprRB gene was able to restore the sporulation wild type phenotype in the ΔnprR-nprRB mutant. Fluorescence assays showed direct binding between NprR and Spo0F, supporting that NprR is a bifunctional protein. To understand how the NprR activation by NprRB could result in two different functions, we studied the molecular recognition mechanism between the signaling peptide and the receptor. Using synthetic variants of NprRB, we found that SSKPDIVG displayed the highest affinity (Kd = 7.19 nM) toward the recombinant NprR and demonstrated that recognition involves conformational selection. We propose that the peptide concentration in the cell controls the oligomerization state of the NprR-NprRB complex for switching between its two functions.
authors.affiliation: Centro de Investigación en Alimentación y Desarrollo A. C., Km 0.6 Carretera a La Victoria, 83304, Hermosillo, Sonora, Mexico.
authors.forename: Rosina
authors.initials: R
authors.lastname: Cabrera
authors.forename: Jorge
authors.initials: J
authors.lastname: Rocha
authors.forename: Víctor
authors.initials: V
authors.lastname: Flores
authors.forename: Luz
authors.initials: L
authors.lastname: Vázquez-Moreno
authors.forename: Gabriel
authors.initials: G
authors.lastname: Guarneros
authors.forename: Gabriela
authors.initials: G
authors.lastname: Olmedo
authors.forename: Adela
authors.initials: A
authors.lastname: Rodríguez-Romero
authors.forename: Mayra
authors.initials: M
authors.lastname: de la Torre
mesh_terms: D001413:Bacillus thuringiensis / Q000235:genetics* / Q000254:growth & development* / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D011485:Protein Binding; D025941:Protein Interaction Mapping; D053038:Quorum Sensing; D015398:Signal Transduction; D013171:Spores, Bacterial / Q000254:growth & development*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; D014157:Transcription Factors