title: N-terminomics identifies Prli42 as a membrane miniprotein conserved in Firmicutes and critical for stressosome activation in Listeria monocytogenes.
abstract: To adapt to changing environments, bacteria have evolved numerous pathways that activate stress response genes. In Gram-positive bacteria, the stressosome, a cytoplasmic complex, relays external cues and activates the sigma B regulon. The stressosome is structurally well-characterized in Bacillus, but how it senses stress remains elusive. Here, we report a genome-wide N-terminomic approach in Listeria that strikingly led to the discovery of 19 internal translation initiation sites and 6 miniproteins, among which one, Prli42, is conserved in Firmicutes. Prli42 is membrane-anchored and interacts with orthologues of Bacillus stressosome components. We reconstituted the Listeria stressosome in vitro and visualized its supramolecular structure by electron microscopy. Analysis of a series of Prli42 mutants demonstrated that Prli42 is important for sigma B activation, bacterial growth following oxidative stress and for survival in macrophages. Taken together, our N-terminonic approach unveiled Prli42 as a long-sought link between stress and the stressosome.
authors.affiliation: Département de Biologie Cellulaire et Infection, Institut Pasteur, Unité des Interactions Bactéries-Cellules, F-75015 Paris, France.|Inserm, U604, F-75015 Paris, France.|INRA, Unité sous-contrat 2020, F-75015 Paris, France.
authors.forename: Francis
authors.initials: F
authors.lastname: Impens
authors.affiliation: Département de Biologie Cellulaire et Infection, Institut Pasteur, Unité des Interactions Bactéries-Cellules, F-75015 Paris, France.|Inserm, U604, F-75015 Paris, France.|INRA, Unité sous-contrat 2020, F-75015 Paris, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Rolhion
authors.affiliation: Département de Biologie Cellulaire et Infection, Institut Pasteur, Unité des Interactions Bactéries-Cellules, F-75015 Paris, France.|Inserm, U604, F-75015 Paris, France.|INRA, Unité sous-contrat 2020, F-75015 Paris, France.
authors.forename: Lilliana
authors.initials: L
authors.lastname: Radoshevich
authors.affiliation: Département de Biologie Cellulaire et Infection, Institut Pasteur, Unité des Interactions Bactéries-Cellules, F-75015 Paris, France.|Inserm, U604, F-75015 Paris, France.|INRA, Unité sous-contrat 2020, F-75015 Paris, France.|Institut Pasteur, Bioinformatics and Biostatistics Hub, C3BI, USR 3756 IP CNRS, Paris, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Bécavin
authors.affiliation: Département de Biologie Cellulaire et Infection, Institut Pasteur, Unité des Interactions Bactéries-Cellules, F-75015 Paris, France.|Inserm, U604, F-75015 Paris, France.|INRA, Unité sous-contrat 2020, F-75015 Paris, France.
authors.forename: Mélodie
authors.initials: M
authors.lastname: Duval
authors.affiliation: Département de Biologie Cellulaire et Infection, Institut Pasteur, Unité des Interactions Bactéries-Cellules, F-75015 Paris, France.|Inserm, U604, F-75015 Paris, France.|INRA, Unité sous-contrat 2020, F-75015 Paris, France.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Mellin
authors.affiliation: Centro Nacional de Biotecnología-Consejo Superior de Investigaciones Científicas (CNB-CSIC), Madrid, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: García Del Portillo
authors.affiliation: Centro Nacional de Biotecnología-Consejo Superior de Investigaciones Científicas (CNB-CSIC), Madrid, Spain.|Departamento de Biología Molecular, Universidad Autónoma de Madrid, Centro de Biología Molecular 'Severo Ochoa' (CBMSO-CSIC), Madrid, Spain.
authors.forename: M Graciela
authors.initials: MG
authors.lastname: Pucciarelli
authors.affiliation: Département de Microbiologie, Institut Pasteur, Unité des Biologie et génétique de la paroi bactérienne, F-75015 Paris, France.|INSERM, Groupe Avenir, F-75015 Paris, France.
authors.forename: Allison H
authors.initials: AH
authors.lastname: Williams
authors.affiliation: Département de Biologie Cellulaire et Infection, Institut Pasteur, Unité des Interactions Bactéries-Cellules, F-75015 Paris, France.|Inserm, U604, F-75015 Paris, France.|INRA, Unité sous-contrat 2020, F-75015 Paris, France.
authors.forename: Pascale
authors.initials: P
authors.lastname: Cossart
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D000068536:Bacillota / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D016680:Genome, Bacterial; D008089:Listeria monocytogenes / Q000235:genetics* / Q000378:metabolism; D008566:Membranes / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000235:genetics; D040901:Proteomics / Q000379:methods; D018085:Regulon / Q000235:genetics; D012808:Sigma Factor / Q000235:genetics; D015398:Signal Transduction; D013312:Stress, Physiological / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D012808:Sigma Factor