151 results on '"Jacob-Dubuisson, Françoise"'
Search Results
2. Structural insights into the signalling mechanisms of two-component systems
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Jacob-Dubuisson, Françoise, Mechaly, Ariel, Betton, Jean-Michel, and Antoine, Rudy
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- 2018
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3. The Crystal Structure of Filamentous Hemagglutinin Secretion Domain and Its Implications for the Two-Partner Secretion Pathway
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Clantin, Bernard, Hodak, Hélène, Willery, Eve, Locht, Camille, Jacob-Dubuisson, Françoise, Villeret, Vincent, and Lipscomb, William N.
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- 2004
4. Development of Pilus Organelle Subassemblies in vitro Depends on Chaperone Uncapping of a Beta Zipper
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Bullitt, Esther, Jones, C. Hal, Striker, Robert, Soto, Gabriel, Jacob-Dubuisson, Francoise, Pinkner, Jerome, Wick, Mary Jo, Makowski, Lee, and Hultgren, Scott J.
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- 1996
5. Two-partner secretion: as simple as it sounds?
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Jacob-Dubuisson, Françoise, Guérin, Jérémy, Baelen, Stéphanie, and Clantin, Bernard
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- 2013
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6. Structural insight into the role of the PAS domain for signal transduction in sensor-kinase BvgS
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Dupré, Elian, Clantin, Bernard, Yuan, Youhua, Lecher, Sophie, Lesne, Elodie, Antoine, Rudy, Villeret, Vincent, Jacob-Dubuisson, Françoise, Jacob-Dubuisson, Françoise, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Institut Pasteur de Lille, and Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS)
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PAS domain ,PAS domain of BvgS Two-component system ,[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,[SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,sensor-kinases ,[SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,virulence regulation ,[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,Bordetella pertussis - Abstract
International audience; The two-component system BvgAS controls the virulence regulon in Bordetella pertussis. BvgS is the prototype of a family of sensor histidine-kinases harboring periplasmic Venus flytrap (VFT) domains. The VFT domains are connected to the cytoplasmic kinase moiety by helical linkers separated by a Per-ARNT-Sim (PAS) domain. Antagonism between the two linkers, as one forms a coiled coil when the other is dynamic and vice versa, regulates BvgS activity. Here we solved the structure of the intervening PAS domain by X-ray
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- 2021
7. Periplasmic domain of the sensor-kinase BvgS reveals a new paradigm for the Venus flytrap mechanism
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Herrou, Julien, Bompard, Coralie, Wintjens, René, Dupré, Elian, Willery, Eve, Villeret, Vincent, Locht, Camille, Antoine, Rudy, and Jacob-Dubuisson, Françoise
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- 2010
8. Structure of the Membrane Protein FhaC: A Member of the Omp85-TpsB Transporter Superfamily
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Clantin, Bernard, Delattre, Anne-Sophie, Rucktooa, Prakash, Saint, Nathalie, Méli, Albano C., Locht, Camille, Jacob-Dubuisson, Françoise, and Villeret, Vincent
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- 2007
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9. Dynamic interplay of membrane-proximal POTRA domain and conserved loop L6 in Omp85 transporter FhaC
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Guérin, Jeremy, Saint, Nathalie, Baud, Catherine, Meli, Albano C., Etienne, Emilien, Locht, Camille, Vezin, Hervé, and Jacob-Dubuisson, Françoise
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- 2015
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10. Outer-Membrane PapC Molecular Usher Discriminately Recognizes Periplasmic Chaperone-Pilus Subunit Complexes
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Dodson, Karen W., Jacob-Dubuisson, Francoise, Striker, Robert T., and Hultgren, Scott J.
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- 1993
11. Conformational dynamics of protein transporter FhaC: large-scale motions of plug helix
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Guérin, Jérémy, Baud, Catherine, Touati, Nadia, Saint, Nathalie, Willery, Eve, Locht, Camille, Vezin, Hervé, and Jacob-Dubuisson, Françoise
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- 2014
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12. Distinct virulence ranges for infection of mice by Bordetella pertussis revealed by engineering of the sensor-kinase BvgS
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Lesne, Elodie, Coutte, Loic, Solans, Luis, Slupek, Stephanie, Debrie, Anne-Sophie, Dhennin, Véronique, Froguel, Philippe, Hot, David, Locht, Camille, Antoine, Rudy, Jacob-Dubuisson, Françoise, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Metabolic functional (epi)genomics and molecular mechanisms involved in type 2 diabetes and related diseases - UMR 8199 - UMR 1283 (EGENODIA (GI3M)), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Metabolic functional (epi)genomics and molecular mechanisms involved in type 2 diabetes and related diseases - UMR 8199 - UMR 1283 (GI3M), and Jacob-Dubuisson, Françoise
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Whooping Cough ,Bordetella ,Molecular biology ,[SDV]Life Sciences [q-bio] ,Gene Expression ,lcsh:Medicine ,Protein Engineering ,Pathology and Laboratory Medicine ,Bordetella pertussis ,DISEASE ,ACTIVATION ,Mice ,Sequencing techniques ,Medicine and Health Sciences ,lcsh:Science ,Lung ,Virulence ,VENUS FLYTRAP ,RNA sequencing ,Animal Models ,Genomics ,INTERMEDIATE PHASE ,Bacterial Pathogens ,[SDV] Life Sciences [q-bio] ,Multidisciplinary Sciences ,RECEPTORS ,Experimental Organism Systems ,Medical Microbiology ,Host-Pathogen Interactions ,BACTERIA ,Science & Technology - Other Topics ,Anatomy ,Pathogens ,Transcriptome Analysis ,Research Article ,RESPIRATORY-INFECTION ,Virulence Factors ,General Science & Technology ,Mouse Models ,Nose ,Research and Analysis Methods ,Microbiology ,TRANSDUCTION ,Model Organisms ,Bacterial Proteins ,MD Multidisciplinary ,Genetics ,Animals ,Animal Models of Disease ,Microbial Pathogens ,Science & Technology ,Sequence Analysis, RNA ,MUTATIONS ,Gene Expression Profiling ,lcsh:R ,Organisms ,Biology and Life Sciences ,Computational Biology ,Gene Expression Regulation, Bacterial ,Genome Analysis ,GENE ,Disease Models, Animal ,Animal Models of Infection ,Molecular biology techniques ,Face ,Mutation ,Animal Studies ,lcsh:Q ,Head ,Transcription Factors - Abstract
International audience; The whooping cough agent Bordetella pertussis coordinately regulates the expression of its virulence factors with the two-component system BvgAS. In laboratory conditions, specific chemical modulators are used to trigger phenotypic modulation of B. pertussis from its default virulent Bvg + phase to avirulent Bvg-or intermediate Bvg i phases, in which no viru-lence factors or only a subset of them are produced, respectively. Whether phenotypic modulation occurs in the host remains unknown. In this work, recombinant B. pertussis strains harboring BvgS variants were tested in a mouse model of infection and analyzed using tran-scriptomic approaches. Recombinant BP-Bvg Δ65, which is in the Bvg i phase by default and can be up-modulated to the Bvg + phase in vitro, could colonize the mouse nose but was rapidly cleared from the lungs, while Bvg +-phase strains colonized both organs for up to four weeks. These results indicated that phenotypic modulation, which might have restored the full virulence capability of BP-Bvg Δ65, does not occur in mice or is temporally or spatially restricted and has no effect in those conditions. Transcriptomic analyses of this and other recombinant Bvg i and Bvg +-phase strains revealed that two distinct ranges of virulence gene expression allow colonization of the mouse nose and lungs, respectively. We also showed that a recombinant strain expressing moderately lower levels of the virulence genes than its wild type parent was as efficient at colonizing both organs. Altogether, genetic modifications of BvgS generate a range of phenotypic phases, which are useful tools to decipher host-pathogen interactions.
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- 2018
13. Current challenges in autotransport and two-partner protein secretion pathways
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Hodak, Hélène and Jacob-Dubuisson, Françoise
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- 2007
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14. Pathogenicity and virulence of Bordetella pertussis and its adaptation to its strictly human host.
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Belcher, Thomas, Dubois, Violaine, Rivera-Millot, Alex, Locht, Camille, and Jacob-Dubuisson, Françoise
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BORDETELLA pertussis ,WHOOPING cough ,HUMAN beings ,ANIMAL models in research - Abstract
The highly contagious whooping cough agent Bordetella pertussis has evolved as a human-restricted pathogen from a progenitor which also gave rise to Bordetella parapertussis and Bordetella bronchiseptica. While the latter colonizes a broad range of mammals and is able to survive in the environment, B. pertussis has lost its ability to survive outside its host through massive genome decay. Instead, it has become a highly successful human pathogen by the acquisition of tightly regulated virulence factors and evolutionary adaptation of its metabolism to its particular niche. By the deployment of an arsenal of highly sophisticated virulence factors it overcomes many of the innate immune defenses. It also interferes with vaccine-induced adaptive immunity by various mechanisms. Here, we review data from invitro, human and animal models to illustrate the mechanisms of adaptation to the human respiratory tract and provide evidence of ongoing evolutionary adaptation as a highly successful human pathogen. [ABSTRACT FROM AUTHOR]
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- 2021
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15. Substrate recognition by the POTRA domains of TpsB transporter FhaC
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Delattre, Anne-Sophie, Saint, Nathalie, Clantin, Bernard, Willery, Eve, Lippens, Guy, Locht, Camille, Villeret, Vincent, and Jacob-Dubuisson, Françoise
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- 2011
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16. Membrane-associated DegP in Bordetella chaperones a repeat-rich secretory protein
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Baud, Catherine, Gutsche, Irina, Willery, Eve, de Paepe, Diane, Drobecq, Hervé, Gilleron, Martine, Locht, Camille, Jamin, Marc, and Jacob-Dubuisson, Françoise
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- 2011
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17. Functional importance of a conserved sequence motif in FhaC, a prototypic member of the TpsB/Omp85 superfamily
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Delattre, Anne-Sophie, Clantin, Bernard, Saint, Nathalie, Locht, Camille, Villeret, Vincent, and Jacob-Dubuisson, Françoise
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- 2010
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18. First structural insights into the TpsB/Omp85 superfamily
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Jacob-Dubuisson, Françoise, Villeret, Vincent, Clantin, Bernard, Delattre, Anne-Sophie, and Saint, Nathalie
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- 2009
19. Secretion signal of the filamentous haemagglutinin, a model two-partner secretion substrate
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Hodak, Hélène, Clantin, Bernard, Willery, Eve, Villeret, Vincent, Locht, Camille, and Jacob-Dubuisson, Françoise
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- 2006
20. Surface anchoring of bacterial subtilisin important for maturation function
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Coutte, Loic, Willery, Eve, Antoine, Rudy, Drobecq, Hervé, Locht, Camille, and Jacob-Dubuisson, Françoise
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- 2003
21. Two-partner secretion in Gram-negative bacteria: a thrifty, specific pathway for large virulence proteins
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Jacob-Dubuisson, Françoise, Locht, Camille, and Antoine, Rudy
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- 2001
22. Evidence that a globular conformation is not compatible with FhaC-mediated secretion of the Bordetella pertussis filamentous haemagglutinin
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Guédin, Sandrine, Willery, Eve, Locht, Camille, and Jacob-Dubuisson, Françoise
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- 1998
23. Amino-terminal maturation of the Bordetella pertussis filamentous haemagglutinin
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Jacob-Dubuisson, Françoise, Buisine, Corinne, Mielcarek, Nathalie, Clément, Eve, Menozzi, Franco D., and Locht, Camille
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- 1996
24. Tetanus toxin fragment C-specific priming by intranasal infection with recombinant Bordetella pertussis
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Reveneau, Nathalie, Alonso, Sylvie, Jacob-Dubuisson, Françoise, Mercenier, Annick, and Locht, Camille
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- 2001
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25. Relationships Between Copper-Related Proteomes and Lifestyles in β Proteobacteria.
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Antoine, Rudy, Rivera-Millot, Alex, Roy, Gauthier, and Jacob-Dubuisson, Françoise
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TRANSITION metals ,COPPER proteins ,COPPER ions ,NUMBERS of species ,NEISSERIA ,PROTEOBACTERIA ,CHARGE exchange - Abstract
Copper is an essential transition metal whose redox properties are used for a variety of enzymatic oxido-reductions and in electron transfer chains. It is also toxic to living beings, and therefore its cellular concentration must be strictly controlled. We have performed in silico analyses of the predicted proteomes of more than one hundred species of β proteobacteria to characterize their copper-related proteomes, including cuproproteins, i.e., proteins with active-site copper ions, copper chaperones, and copper-homeostasis systems. Copper-related proteomes represent between 0 and 1.48% of the total proteomes of β proteobacteria. The numbers of cuproproteins are globally proportional to the proteome sizes in all phylogenetic groups and strongly linked to aerobic respiration. In contrast, environmental bacteria have considerably larger proportions of copper-homeostasis systems than the other groups of bacteria, irrespective of their proteome sizes. Evolution toward commensalism, obligate, host-restricted pathogenesis or symbiosis is globally reflected in the loss of copper-homeostasis systems. In endosymbionts, defense systems and copper chaperones have disappeared, whereas residual cuproenzymes are electron transfer proteins for aerobic respiration. Lifestyle is thus a major determinant of the size and composition of the copper-related proteome, and it is particularly reflected in systems involved in copper homeostasis. Analyses of the copper-related proteomes of a number of species belonging to the Burkholderia , Bordetella , and Neisseria genera indicates that commensals are in the process of shedding their copper-homeostasis systems and chaperones to greater extents yet than pathogens. [ABSTRACT FROM AUTHOR]
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- 2019
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26. Two-partner Secretion of Gram-negative Bacteria: A SINGLE β-BARREL PROTEIN ENABLES TRANSPORT ACROSS THE OUTER MEMBRANE*
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Fan, Enguo, Fiedler, Silke, Jacob-Dubuisson, Françoise, and Müller, Matthias
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Protein Transport ,Cell-Free System ,Membrane Biology ,macromolecular substances ,respiratory system ,bacterial infections and mycoses ,Carrier Proteins ,Bacterial Secretion Systems ,Bordetella pertussis ,respiratory tract diseases ,Bacterial Outer Membrane Proteins - Abstract
The mechanisms of protein secretion by pathogenic bacteria remain poorly understood. In gram-negative bacteria, the two-partner secretion pathway exports large, mostly virulence-related "TpsA" proteins across the outer membrane via their dedicated "TpsB" transporters. TpsB transporters belong to the ubiquitous Omp85 superfamily, whose members are involved in protein translocation across, or integration into, cellular membranes. The filamentous hemagglutinin/FhaC pair of Bordetella pertussis is a model two-partner secretion system. We have reconstituted the TpsB transporter FhaC into proteoliposomes and demonstrate that FhaC is the sole outer membrane protein required for translocation of its cognate TpsA protein. This is the first in vitro system for analyzing protein secretion across the outer membrane of gram-negative bacteria. Our data also provide clear evidence for the protein translocation function of Omp85 transporters.
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- 2011
27. Characterization of a Bvg-regulated fatty acid methyl-transferase in Bordetella pertussis.
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Rivera-Millot, Alex, Lesne, Elodie, Solans, Luis, Coutte, Loic, Bertrand-Michel, Justine, Froguel, Philippe, Dhennin, Véronique, Hot, David, Locht, Camille, Antoine, Rudy, and Jacob-Dubuisson, Françoise
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BORDETELLA pertussis ,METHYLTRANSFERASES ,WHOOPING cough ,FATTY acids ,MICROBIAL virulence ,CELLULAR signal transduction - Abstract
The whooping cough agent Bordetella pertussis controls the expression of its large virulence regulon in a coordinated manner through the two-component signal transduction system BvgAS. In addition to the genes coding for bona fide virulence factors, the Bvg regulon comprises genes of unknown function. In this work, we characterized a new Bvg-activated gene called BP2936. Homologs of BP2936 are found in other pathogenic Bordetellae and in several other species, including plant pathogens and environmental bacteria. We showed that the gene product of BP2936 is a membrane-associated methyl-transferase of free fatty acids. We thus propose to name it FmtB, for atty acid ethyl-ransferase of ordetella. The role of this protein was tested in cellular and animal models of infection, but the loss of BP2936 did not appear to affect host-pathogen interactions in those assays. The high level of conservation of BP2936 among B. pertussis isolates nevertheless argues that it probably plays a role in the life cycle of this pathogen. [ABSTRACT FROM AUTHOR]
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- 2017
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28. Identification of a Ligand Binding Pocket in LdtR from Liberibacter asiaticus.
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Pagliai, Fernando A., Gonzalez, Claudio F., Lorca, Graciela L., Jacob-Dubuisson, Françoise, Stevenson, Clare Elizabeth Mary, and Rubio, Vicente
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FREMYELLA diplosiphon ,MORPHOGENESIS ,CYANOBACTERIA - Abstract
LdtR is a transcriptional activator involved in the regulation of a putative L,D transpeptidase in Liberibacter asiaticus, an unculturable pathogen and one of the causative agents of Huanglongbing disease. Using small molecule screens we identified benzbromarone as an inhibitor of LdtR activity, which was confirmed using in vivo and in vitro assays. Based on these previous results, the objective of this work was to identify the LdtR ligand binding pocket and characterize its interactions with benzbromarone. A structural model of LdtR was constructed and the molecular interactions with the ligand were predicted using the SwissDock interface. Using site-directed mutagenesis, these residues were changed to alanine. Electrophoretic mobility shift assays, thermal denaturation, isothermal titration calorimetry experiments, and in vivo assays were used to identify residues T43, L61, and F64 in the Benz1 pocket of LdtR as the amino acids most likely involved in the binding to benzbromarone. These results provide new information on the binding mechanism of LdtR to a modulatory molecule and provide a blue print for the design of therapeutics for other members of the MarR family of transcriptional regulators involved in pathogenicity. [ABSTRACT FROM AUTHOR]
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- 2015
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29. Virulence Regulation with Venus Flytrap Domains: Structure and Function of the Periplasmic Moiety of the Sensor-Kinase BvgS.
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Dupré, Elian, Herrou, Julien, Lensink, Marc F., Wintjens, René, Vagin, Alexey, Lebedev, Andrey, Crosson, Sean, Villeret, Vincent, Locht, Camille, Antoine, Rudy, and Jacob-Dubuisson, Françoise
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BORDETELLA pertussis ,VENUS'S flytrap ,MUTAGENESIS ,MICROBIAL virulence ,KINASES - Abstract
Two-component systems (TCS) represent major signal-transduction pathways for adaptation to environmental conditions, and regulate many aspects of bacterial physiology. In the whooping cough agent Bordetella pertussis, the TCS BvgAS controls the virulence regulon, and is therefore critical for pathogenicity. BvgS is a prototypical TCS sensor-kinase with tandem periplasmic enus lyrap (VFT) domains. VFT are bi-lobed domains that typically close around specific ligands using clamshell motions. We report the X-ray structure of the periplasmic moiety of BvgS, an intricate homodimer with a novel architecture. By combining site-directed mutagenesis, functional analyses and molecular modeling, we show that the conformation of the periplasmic moiety determines the state of BvgS activity. The intertwined structure of the periplasmic portion and the different conformation and dynamics of its mobile, membrane-distal VFT1 domains, and closed, membrane-proximal VFT2 domains, exert a conformational strain onto the transmembrane helices, which sets the cytoplasmic moiety in a kinase-on state by default corresponding to the virulent phase of the bacterium. Signaling the presence of negative signals perceived by the periplasmic domains implies a shift of BvgS to a distinct state of conformation and activity, corresponding to the avirulent phase. The response to negative modulation depends on the integrity of the periplasmic dimer, indicating that the shift to the kinase-off state implies a concerted conformational transition. This work lays the bases to understand virulence regulation in Bordetella. As homologous sensor-kinases control virulence features of diverse bacterial pathogens, the BvgS structure and mechanism may pave the way for new modes of targeted therapeutic interventions. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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30. Translocation path of a substrate protein through its Omp85 transporter.
- Author
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Baud, Catherine, Guérin, Jérémy, Petit, Emmanuelle, Lesne, Elodie, Dupré, Elian, Locht, Camille, and Jacob-Dubuisson, Françoise
- Abstract
TpsB proteins are Omp85 superfamily members that mediate protein translocation across the outer membrane of Gram-negative bacteria. Omp85 transporters are composed of N-terminal POTRA domains and a C-terminal transmembrane b-barrel. In this work, we track the in vivo secretion path of the Bordetella pertussis filamentous haemagglutinin (FHA), the substrate of the model TpsB transporter FhaC, using site-specific crosslinking. The conserved secretion domain of FHA interacts with the POTRA domains, specific extracellular loops and strands of FhaC and the inner b-barrel surface. The interaction map indicates a funnel-like pathway, with conformationally flexible FHA entering the channel in a non-exclusive manner and exiting along a four-stranded b-sheet at the surface of the FhaC barrel. This sheet of FhaC guides the secretion domain of FHA along discrete steps of translocation and folding. This work demonstrates that the Omp85 barrel serves as a channel for translocation of substrate proteins. [ABSTRACT FROM AUTHOR]
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- 2014
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31. Sequential Unfolding of Beta Helical Protein by Single-Molecule Atomic Force Microscopy.
- Author
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Alsteens, David, Martinez, Nicolas, Jamin, Marc, and Jacob-Dubuisson, Françoise
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PROTEIN folding ,ATOMIC force microscopy ,EXTRACELLULAR matrix proteins ,GRAM-negative bacteria ,MOLECULAR dynamics ,BORDETELLA pertussis ,CARRIER proteins - Abstract
The parallel βhelix is a common fold among extracellular proteins, however its mechanical properties remain unexplored. In Gram-negative bacteria, extracellular proteins of diverse functions of the large ‘TpsA’ family all fold into long βhelices. Here, single-molecule atomic force microscopy and steered molecular dynamics simulations were combined to investigate the mechanical properties of a prototypic TpsA protein, FHA, the major adhesin of Bordetella pertussis. Strong extension forces were required to fully unfold this highly repetitive protein, and unfolding occurred along a stepwise, hierarchical process. Our analyses showed that the extremities of the βhelix unfold early, while central regions of the helix are more resistant to mechanical unfolding. In particular, a mechanically resistant subdomain conserved among TpsA proteins and critical for secretion was identified. This nucleus harbors structural elements packed against the βhelix that might contribute to stabilizing the N-terminal region of FHA. Hierarchical unfolding of the βhelix in response to a mechanical stress may maintain β-helical portions that can serve as templates for regaining the native structure after stress. The mechanical properties uncovered here might apply to many proteins with β-helical or related folds, both in prokaryotes and in eukaryotes, and play key roles in their structural integrity and functions. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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32. Characterization of the PAS domain in the sensor-kinase BvgS: mechanical role in signal transmission.
- Author
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Dupré, Elian, Wohlkonig, Alexandre, Herrou, Julien, Locht, Camille, Jacob-Dubuisson, Françoise, and Antoine, Rudy
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KINASES ,AUTOPHOSPHORYLATION ,PHOSPHORYL group ,BORDETELLA pertussis ,VENUS'S flytrap ,MUTAGENESIS ,POLYALUMINUM sulfate - Abstract
Background: In bacteria, signal-transduction two-component systems are major players for adaptation to environmental stimuli. The perception of a chemical or physical signal by a sensor-kinase triggers its autophosphorylation. The phosphoryl group is then transferred to the cognate response regulator, which mediates the appropriate adaptive response. Virulence of the whooping cough agent Bordetella pertussis is controlled by the two-component system BvgAS. Atypically, the sensor-kinase BvgS is active without specific stimuli at 37°C in laboratory conditions and is inactivated by the addition of negative chemical modulators. The structure of BvgS is complex, with two tandem periplasmic Venus flytrap domains and a cytoplasmic PAS domain that precedes the kinase domain, which is followed by additional phosphotransfer domains. PAS domains are small, ubiquitous sensing or regulatory domains. The function of the PAS domain in BvgS remains unknown. Results: We showed that recombinant BvgS PAS proteins form dimers that are stabilized by α helical regions flanking the PAS core. A structural model of the PAS domain dimer was built and probed by site-directed mutagenesis and by biochemical and functional analyses. Although we found no ligands for the PAS domain cavity, its integrity is required for signaling. We also showed that the structural stability of the PAS core and its proper coupling to its flanking N- and C-terminal α helices are crucial for BvgS activity. Conclusions: We propose that a major function of the BvgS PAS domain is to maintain conformational signals arising from mechanical strain generated by the periplasmic domain. The tight structure of the PAS core and its connections with the upstream and downstream helices ensure signaling to the kinase domain, which determines BvgS activity. Many mild substitutions that map to the PAS domain keep BvgS active but make it unresponsive to negative modulators, supporting that modulation increases conformational strain in the protein. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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33. Molecular Evolution of the Two-Component System BvgAS Involved in Virulence Regulation in Bordetella.
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Herrou, Julien, Debrie, Anne-Sophie, Willery, Eve, Renaud-Mongénie, Geneviève, Locht, Camille, Mooi, Frits, Jacob-Dubuisson, Françoise, and Antoine, Rudy
- Subjects
MICROBIAL virulence ,BORDETELLA pertussis ,WHOOPING cough ,GENETIC regulation ,RESPIRATORY diseases ,GRAM-negative bacterial diseases ,FOCAL adhesion kinase ,MICROBIAL genetics ,MICROBIOLOGY - Abstract
The whooping cough agent Bordetella pertussis is closely related to Bordetella bronchiseptica, which is responsible for chronic respiratory infections in various mammals and is occasionally found in humans, and to Bordetella parapertussis, one lineage of which causes mild whooping cough in humans and the other ovine respiratory infections. All three species produce similar sets of virulence factors that are co-regulated by the two-component system BvgAS. We characterized the molecular diversity of BvgAS in Bordetella by sequencing the two genes from a large number of diverse isolates. The response regulator BvgA is virtually invariant, indicating strong functional constraints. In contrast, the multi-domain sensor kinase BvgS has evolved into two different types. The pertussis type is found in B. pertussis and in a lineage of essentially human-associated B. bronchiseptica, while the bronchiseptica type is associated with the majority of B. bronchiseptica and both ovine and human B. parapertussis. BvgS is monomorphic in B. pertussis, suggesting optimal adaptation or a recent population bottleneck. The degree of diversity of the bronchiseptica type BvgS is markedly different between domains, indicating distinct evolutionary pressures. Thus, absolute conservation of the putative solute-binding cavities of the two periplasmic Venus Fly Trap (VFT) domains suggests that common signals are perceived in all three species, while the external surfaces of these domains vary more extensively. Co-evolution of the surfaces of the two VFT domains in each type and domain swapping experiments indicate that signal transduction in the periplasmic region may be type-specific. The two distinct evolutionary solutions for BvgS confirm that B. pertussis has emerged from a specific B. bronchiseptica lineage. The invariant regions of BvgS point to essential parts for its molecular mechanism, while the variable regions may indicate adaptations to different lifestyles. The repertoire of BvgS sequences will pave the way for functional analyses of this prototypic system. [ABSTRACT FROM AUTHOR]
- Published
- 2009
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34. Influence of the passenger domain of a model autotransporter on the properties of its translocator domain.
- Author
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Dé, Emmanuelle, Saint, Nathalie, Glinel, Karine, Meli, Albano C., Lévy, Daniel, and Jacob-Dubuisson, Françoise
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BIOMOLECULES ,BACTERIA ,PROTEINS ,AMINO acids ,LIGHT scattering - Abstract
Autotransporters are a superfamily of proteins secreted by Gram-negative bacteria including many virulence factors. They are modular proteins composed of an N-terminal signal peptide, a surface-exposed 'passenger' domain carrying the activity of the protein, and a C-terminal 'translocator' domain composed of an α-helical linker region and a transmembrane β-barrel. The translocator domain plays an essential role for the secretion of the passenger domain across the outer membrane; however, the mechanism of autotransport remains poorly understood. The whooping cough agent Bordetella pertussis produces an autotransporter serine-protease, SphB1, which is involved in the maturation of an adhesin at the bacterial surface. SphB1 also mediates the proteolytic maturation of its own precursor. We used SphB1 as a model autotransporter and performed the first comparisons of the biochemical and biophysical properties of an isolated translocator domain with those of the same domain preceded by the C-terminal moiety of its natural passenger. By using cross-linking and dynamic light scattering, we provide evidence that the passenger domain promotes the auto-association of SphB1, although these interactions appear rather labile. Electrophysiological studies revealed that the passenger domain of the autotransporter appears to maintain the translocator channel in a low-conductance conformation, most likely by stabilizing the α-helix inside the pore. That the passenger may significantly influence AT physicochemical properties is likely to be relevant for the in vivo maturation and stability of AT proteins. [ABSTRACT FROM AUTHOR]
- Published
- 2008
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35. Structural analysis of Bordetella pertussis BugE solute receptor in a bound conformation.
- Author
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Huvent, Isabelle, Locht, Camille, Belrhali, Hassan, Antoine, Rudy, Bompard, Coralie, Jacob-Dubuisson, Françoise, and Villeret, Vincent
- Subjects
BORDETELLA pertussis ,PROTEINS ,AMINO acids ,CRYSTALLIZATION ,LIGAND binding (Biochemistry) - Abstract
The Bug proteins form a large family of periplasmic solute-binding receptors present in a number of bacterial species. Here, the crystal structure of Bordetella pertussis BugE, a member of the Bug family coded by the gene BP0250, is reported. It adopts the Venus flytrap architecture of periplasmic binding proteins, with two domains separated by a deep cleft. BugE has a bound ligand, identified as a glutamate. The structure of B. pertussis BugD, which is an aspartic acid transporter, has recently been reported. These structures reveal high conservation of the Bug architecture, despite limited sequence identity. They share a common carboxylate-binding motif defined by two strand–β-turn–α-helix motifs, also involving two water molecules to bridge the carboxylate O atoms to the protein. The two water molecules are hydrogen bonded to a common main-chain carbonyl group. Although the features of the carboxylate-binding motif are totally conserved, the ligand in BugE is bound by its side-chain carboxylate group rather than by its α-carboxylate as in BugD. This specific ligand-binding motif is highly conserved in Bug proteins and the BugE structure suggests that the cavity of the Bug proteins might also accommodate carboxylated solutes other than amino acids. The vast expansion of the Bug family in several bacterial genera is likely to be explained by the possible diversity of ligands. No charged residues are involved in glutamate binding by BugE, unlike what has been described for all glutamate receptors reported so far. [ABSTRACT FROM AUTHOR]
- Published
- 2006
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36. Membrane Targeting of a Bacterial Virulence Factor Harbouring an Extended Signal Peptide.
- Author
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Chevalier, Nina, Moser, Michael, Koch, Hans-Georg, Schimz, Karl-Ludwig, Willery, Eve, Locht, Camille, Jacob-Dubuisson, Françoise, and Müller, Matthias
- Subjects
GRAM-negative bacterial diseases ,RESPIRATORY infections ,BIOLOGICAL transport ,ENTEROBACTERIACEAE ,ESCHERICHIA coli ,MEMBRANE proteins ,BIOLOGICAL membranes - Abstract
Filamentous haemagglutinin (FHA) is the major adhesin of Bordetella pertussis, the whooping cough agent. FHA is synthesized as a 367-kDa precursor harbouring a remarkably long signal peptide with an N-terminal extension that is conserved among related virulence proteins. FHA is secreted via the two-partner secretion pathway that involves transport across the outer membrane by a cognate transporter protein. Here we have analyzed the mechanism by which FHA is targeted to, and translocated across, the inner membrane. Studies were performed both in vitro using Escherichia coli inside-out inner membrane vesicles and in vivo by pulse-chase labelling of Bordetella pertussis cells. The data collectively indicate that like classical periplasmic and outer membrane proteins, FHA requires SecA and SecB for its export through the SecYEG translocon in the inner membrane. Although short nascent chains of FHA were found to cross-link to signal recognition particle (SRP), we did not obtain indication for an SRP-dependent, co-translational membrane targeting provoked by the FHA signal sequence. Our results rule out that the extended signal peptide of FHA determines a specific mode of membrane targeting but rather suggest that it might influence the export rate at the inner membrane. Copyright © 2004 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]
- Published
- 2005
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37. Protein secretion through autotransporter and two-partner pathways
- Author
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Jacob-Dubuisson, Françoise, Fernandez, Rachel, and Coutte, Loic
- Subjects
- *
PROTEINS , *BIOMOLECULES , *BACTERIA , *PROKARYOTES - Abstract
Two distinct protein secretion pathways, the autotransporter (AT) and the two-partner secretion (TPS) pathways are characterized by their apparent simplicity. Both are devoted to the translocation across the outer membrane of mostly large proteins or protein domains. As implied by their name, AT proteins contain their own transporter domain, covalently attached to the C-terminal extremity of the secreted passenger domain, while TPS systems are composed of two separate proteins, with TpsA being the secreted protein and TpsB its specific transporter. In both pathways, the secreted proteins are exported in a Sec-dependent manner across the inner membrane, after which they cross the outer membrane with the help of their cognate transporters. The AT translocator domains and the TpsB proteins constitute distinct families of protein-translocating, outer membrane porins of Gram-negative bacteria. Both types of transporters insert into the outer membrane as β-barrel proteins possibly forming oligomeric pores in the case of AT and serve as conduits for their cognate secreted proteins or domains across the outer membrane. Translocation appears to be folding-sensitive in both pathways, indicating that AT passenger domains and TpsA proteins cross the periplasm and the outer membrane in non-native conformations and fold progressively at the cell surface.A major difference between AT and TPS pathways arises from the manner by which specificity is established between the secreted protein and its transporter. In AT, the covalent link between the passenger and the translocator domains ensures the translocation of the former without the need for a specific molecular recognition between the two modules. In contrast, the TPS pathway has solved the question of specific recognition between the TpsA proteins and their transporters by the addition to the TpsA proteins of an N-proximal module, the conserved TPS domain, which represents a hallmark of the TPS pathway. [Copyright &y& Elsevier]
- Published
- 2004
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38. Overrepresentation of a Gene Family Encoding Extracytoplasmic Solute Receptors in Bordetella.
- Author
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Antoine, Rudy, Jacob-Dubuisson, Françoise, Drobecq, Hervé, Willery, Eve, Lesjean, Sarah, and Locht, Camille
- Subjects
- *
BORDETELLA pertussis , *GENE expression - Abstract
A family of genes that are likely to encode extracytoplasmic solute receptors is strongly overrepresented in several β-proteobacteria, including Bordetella pertussis. This gene family, of which members have been called bug genes, contains some examples that are contained within polycistronic operons coding for tripartite uptake transporters of the TTT family, while the vast majority are "orphan" genes. Proteomic and functional analyses demonstrated that several of these genes are expressed in B. pertussis, and one is involved in citrate uptake. The bug genes probably form an ancient family that has been subjected to a large expansion in a restricted phylogenic group. [ABSTRACT FROM AUTHOR]
- Published
- 2003
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39. Subtilisin-like autotransporter serves as maturation protease in a bacterial secretion pathway.
- Author
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Coutte, Loic, Antoine, Rudy, Drobecq, Herve, Locht, Camille, and Jacob-Dubuisson, Françoise
- Subjects
PROTEINS ,SUBTILISINS ,BACTERIA ,MOLECULAR chaperones ,MICROBIAL enzymes ,BORDETELLA pertussis - Abstract
Proteins of Gram-negative bacteria destined to the extracellular milieu must cross the two cellular membranes and then fold at the appropriate time and place. The synthesis of a precursor may be a strategy to maintain secretion competence white preventing aggregation or premature folding (especially for large proteins). The secretion of 230 kDa filamentous haemagglutinin (FHA) of Bordetella pertussis requires the synthesis and the maturation of a 367 kDa precursor that undergoes the proteolytic removal of its ∼130 kDa C-terminal intramolecular chaperone domain. We have identified a specific protease, SphB1, responsible for the timely maturation of the precursor FhaB, which allows for extracellular release of FHA. SphB1 is a large exported protein with a subtilisin-like domain and a C-terminal domain typical of bacterial autotransporters. SphB1 is the first described subtilisin-like protein that serves as a specialized maturation protease in a secretion pathway of Gram-negative bacteria. This is reminiscent of pro-protein convertases of eukaryotic cells. [ABSTRACT FROM AUTHOR]
- Published
- 2001
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40. N-terminal characterization of the Bordetella pertussis filamentous haemagglutinin.
- Author
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Lambert-Buisine, Corinne, Willery, Eve, Locht, Camille, and Jacob-Dubuisson, Françoise
- Subjects
HEMAGGLUTININ ,BORDETELLA pertussis - Abstract
The major adhesin of Bordetella pertussis , filamentous haemagglutinin (FHA), is produced and secreted at high levels by the bacterium. Mature FHA derives from a large precursor, FhaB, that undergoes several post-translational maturations. In this work, we demonstrate by site-directed mutagenesis that the N-terminal signal peptide of FHA is composed of 71 amino acids, including a 22-residue-long ‘N-terminal extension’ sequence. This sequence, although highly conserved in various other secretory proteins, does not appear to play an essential part in FHA secretion, as shown by deletion mutagenesis. The entire N-terminal signal region of FhaB is removed in the course of secretion by proteolytic cleavage at a site that corresponds to a Lep signal peptidase recognition sequence. After this maturation, the N-terminal glutamine residue is modified to a pyroglutamate residue. This modification is not crucial for heparin binding, haemagglutination or secretion. Interestingly, however, the modification is absent from Escherichia coli secreted FHA derivatives. In addition, it is dependent in B. pertussis on the presence of all three cysteines contained in the signal peptide of FhaB. These observations suggest that it does not occur spontaneously but perhaps requires a specific enzymatic machinery. [ABSTRACT FROM AUTHOR]
- Published
- 1998
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- View/download PDF
41. Crystallization and preliminary X-ray diffraction analysis of two extracytoplasmic solute receptors of the DctP family from Bordetella pertussis.
- Author
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Rucktooa, Prakash, Huvent, Isabelle, Antoine, Rudy, Lecher, Sophie, Jacob-Dubuisson, Françoise, Villeret, Vincent, and Bompard, Coralie
- Subjects
BORDETELLA pertussis ,PROTEINS ,X-ray diffraction ,CRYSTALLIZATION ,CYTOPLASM ,CARRIER proteins - Abstract
DctP6 and DctP7 are two Bordetella pertussis proteins which belong to the extracytoplasmic solute receptors (ESR) superfamily. ESRs are involved in the transport of substrates from the periplasm to the cytosol of Gram-negative bacteria. DctP6 and DctP7 have been crystallized and diffraction data were collected using a synchrotron-radiation source. DctP6 crystallized in space group P4
1 21 2, with unit-cell parameters a = 108.39, b = 108.39, c = 63.09 Å, while selenomethionyl-derivatized DctP7 crystallized in space group P21 21 21 , with unit-cell parameters a = 64.87, b = 149.83, c = 170.65 Å. The three-dimensional structure of DctP7 will be determined by single-wavelength anomalous diffraction, while the DctP6 structure will be solved by molecular-replacement methods. [ABSTRACT FROM AUTHOR]- Published
- 2006
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- View/download PDF
42. Autotransporter proteins, evolution and redefining protein secretion: Response
- Author
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Jacob-Dubuisson, Françoise, Antoine, Rudy, and Locht, Camille
- Published
- 2000
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- View/download PDF
43. Conserved Omp85 lid-lock structure and substrate recognition in FhaC.
- Author
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Maier, Timm, Clantin, Bernard, Gruss, Fabian, Dewitte, Frédérique, Delattre, Anne-Sophie, Jacob-Dubuisson, Françoise, Hiller, Sebastian, and Villeret, Vincent
- Published
- 2015
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44. Versatile in vitro system to study translocation and functional integration of bacterial outer membrane proteins.
- Author
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Norell, Derrick, Heuck, Alexander, Tran-Thi, Thuy-Anh, Götzke, Hansjörg, Jacob-Dubuisson, Françoise, Clausen, Tim, Daley, Daniel O., Braun, Volkmar, Müller, Matthias, and Fan, Enguo
- Published
- 2014
- Full Text
- View/download PDF
45. A novel secretion apparatus for the assembly of adhesive bacterial pili
- Author
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Jacob-Dubuisson, Françoise, Kuehn, Meta, and Hultgren, Scott J.
- Published
- 1993
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- View/download PDF
46. Two-partner Secretion of Gram-negative Bacteria A SINGLEβ-BARREL PROTEIN ENABLES TRANSPORT ACROSS THE OUTER MEMBRANE.
- Author
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Fan, Enguo, Fiedler, Silke, Jacob-Dubuisson, Françoise, and Müller, Matthias
- Subjects
- *
GRAM-negative bacteria , *MICROBIAL virulence , *PROTEINS , *SECRETION , *PATHOGENIC bacteria - Abstract
The mechanisms of protein secretion by pathogenic bacteria remain poorly understood. In Gram-negative bacteria, the twopartner secretion pathway exports large, mostly virulence-related "TpsA" proteins across the outer membrane via their dedicated "TpsB" transporters. TpsB transporters belong to the ubiquitous Omp85 superfamily, whose members are involved in protein translocation across, or integration into, cellular membranes. The filamentous hemagglutinin/FhaC pair of Bordetella pertussis is a model two-partner secretion system. We have reconstituted the TpsB transporter FhaC into proteoliposomes and demonstrate that FhaC is the sole outer membrane protein required for translocation of its cognate TpsA protein. This is the first in vitro system for analyzing protein secretion across the outer membrane of Gram-negative bacteria. Our data also provide clear evidence for the protein translocation function of Omp85 transporters. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
47. Posttranscriptional Regulation by Copper with a New Upstream Open Reading Frame
- Author
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Gauthier Roy, Rudy Antoine, Annie Schwartz, Stéphanie Slupek, Alex Rivera-Millot, Marc Boudvillain, Françoise Jacob-Dubuisson, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Jacob-Dubuisson, Françoise
- Subjects
upstream ORF ,copper homeostasis ,posttranscriptional regulation ,Gene Expression Regulation, Bacterial ,Microbiology ,[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,Bordetella pertussis ,Open Reading Frames ,Bacterial Proteins ,Gene Expression Regulation ,Virology ,DUF2946 family ,[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,Operon ,[SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,[SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,Ribosomes ,Copper - Abstract
International audience; Copper is essential to most living beings but also highly toxic and as such is an important player at the host-pathogen interface. Bacteria have thus developed homeostatic mechanisms to tightly control its intracellular concentration. Known Cu export and import systems are under transcriptional control, whereas post-transcriptional regulatory mechanisms are yet to be characterized. We identified a three-gene operon, bp2923-bfrG-bp2921, down-regulated by copper and notably encoding a TonB-dependent transporter in Bordetella pertussis. We show here that the protein encoded by the first gene, which is a member of the DUF2946 protein family, represents a new type of upstream Open Reading Frame (uORF) involved in post-transcriptional regulation of the downstream genes. In the absence of copper, the entire operon is transcribed and translated. Perception of copper by the nascent bp2923-coded protein via its conserved CXXC motif triggers Rho-dependent transcription termination between the first and second genes by relieving translation arrest on a conserved C-terminal RAPP motif. Homologs of bp2923 are widespread in bacterial genomes, where they head operons predicted to participate in copper homeostasis. This work has thus unveiled a new mode of genetic regulation by a transition metal and identified a regulatory function for a member of an uncharacterized family of bacterial proteins that we have named CruR, for copper-responsive upstream regulator.
- Published
- 2022
48. Large-scale conformational changes of FhaC provide insights into the two-partner secretion mechanism
- Author
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S. Lecher, Robert Schneider, G. Rouaut, Hervé Vezin, Françoise Jacob-Dubuisson, L. Müggenburg, Oscar Hernandez-Alba, Frank Sobott, E. Del Nero, Sarah Cianférani, J. Guerin, G. Sicoli, Steve Hessmann, Albert Konijnenberg, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement - UMR 8516 [LASIRE], University of Antwerp [UA], Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 [CIIL], Laboratoire de Spectrométrie de Masse BioOrganique [Strasbourg] [LSMBO], Facteurs de Risque et Déterminants Moléculaires des Maladies liées au Vieillissement - U 1167 [RID-AGE], University of Leeds, Université de Lille, CNRS, Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement (LASIRE) - UMR 8516, Centre d'Infection et d'Immunité de Lille (CIIL) - U1019 - UMR 9017, Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement - UMR 8516 (LASIRE), Institut de Chimie du CNRS (INC)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), University of Antwerp (UA), Laboratoire de Spectrométrie de Masse BioOrganique [Strasbourg] (LSMBO), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Infrastructure Nationale de Protéomique, FR2048 ProFI, Biologie Structurale Intégrative (ERL 9002 - BSI ), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Facteurs de Risque et Déterminants Moléculaires des Maladies liées au Vieillissement - U 1167 (RID-AGE), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), CHU Lille, Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), and Jacob-Dubuisson, Françoise
- Subjects
[SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,Beta hairpin ,Omp85 superfamily ,Gramnegative bacteria ,010402 general chemistry ,two-partner secretion system ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,Biochemistry ,01 natural sciences ,outer membrane protein ,03 medical and health sciences ,[CHIM]Chemical Sciences ,Secretion ,Lipid bilayer ,Molecular Biology ,Biology ,030304 developmental biology ,mass spectrometry ,0303 health sciences ,[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,Chemistry ,Periplasmic space ,[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,Transmembrane protein ,NMR ,0104 chemical sciences ,Transport protein ,[CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry ,[CHIM.THEO] Chemical Sciences/Theoretical and/or physical chemistry ,Beta barrel ,protein dynamics ,Biophysics ,EPR ,[SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,Bacterial outer membrane - Abstract
The Two-Partner secretion pathway mediates protein transport across the outer membrane of Gram-negative bacteria. TpsB transporters belong to the Omp85 superfamily, whose members catalyze protein insertion into, or translocation across membranes without external energy sources. They are composed of a transmembrane β barrel preceded by two periplasmic POTRA domains that bind the incoming protein substrate. Here we used an integrative approach combining in vivo assays, mass spectrometry, nuclear magnetic resonance and electron paramagnetic resonance techniques suitable to detect minor states in heterogeneous populations, to explore transient conformers of the TpsB transporter FhaC. This revealed substantial, spontaneous conformational changes with a portion of the POTRA2 domain coming close to the lipid bilayer and surface loops. Specifically, the amphipathic β hairpin immediately preceding the first barrel strand can insert into the β barrel. We propose that these motions enlarge the channel and hoist the substrate into it for secretion. An anchor region at the interface of the β barrel and the POTRA2 domain stabilizes the transporter in the course of secretion. Our data propose a solution to the conundrum how these transporters mediate protein secretion without the need for cofactors, by utilizing intrinsic protein dynamics.
- Published
- 2021
49. Pathogenicity and virulence of Bordetella pertussis and its adaptation to its strictly human host
- Author
-
Alex Rivera-Millot, Camille Locht, Françoise Jacob-Dubuisson, Thomas Belcher, Violaine Dubois, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), and Jacob-Dubuisson, Françoise
- Subjects
Microbiology (medical) ,Bordetella pertussis ,Bordetella ,Immunology ,virulence factors ,Virulence ,Human pathogen ,Infectious and parasitic diseases ,RC109-216 ,Microbiology ,Bordetella parapertussis ,03 medical and health sciences ,Pertussis ,evolution ,Pathogen ,innate immunity ,030304 developmental biology ,Genetics ,0303 health sciences ,Bordetella bronchiseptica ,biology ,030306 microbiology ,adaptive immunity ,biology.organism_classification ,Acquired immune system ,[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,3. Good health ,Infectious Diseases ,Parasitology ,[SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,metabolism - Abstract
International audience; The highly contagious whooping cough agent Bordetella pertussis has evolved as a humanrestricted pathogen from a progenitor which also gave rise to Bordetella parapertussis and Bordetella bronchiseptica. While the latter colonizes a broad range of mammals and is able to survive in the environment, B. pertussis has lost its ability to survive outside its host through massive genome decay. Instead, it has become a highly successful human pathogen by the acquisition of tightly regulated virulence factors and evolutionary adaptation of its metabolism to its particular niche. By the deployment of an arsenal of highly sophisticated virulence factors it overcomes many of the innate immune defenses. It also interferes with vaccine-induced adaptive immunity by various mechanisms. Here, we review data from in vitro, human and animal models to illustrate the mechanisms of adaptation to the human respiratory tract and provide evidence of ongoing evolutionary adaptation as a highly successful human pathogen.
- Published
- 2021
50. Structural insight into the role of the PAS domainfor signal transduction in sensor-kinase BvgS
- Author
-
Youhua Yuan, Elian Dupré, Elodie Lesne, Françoise Jacob-Dubuisson, Rudy Antoine, Bernard Clantin, Vincent Villeret, Sophie Lecher, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Facteurs de Risque et Déterminants Moléculaires des Maladies liées au Vieillissement - U 1167 (RID-AGE), Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, This work was initiated with the grant ANR-13-BSV8-0002-01 to FJD and then pursued with the financial support of Inserm and the University of Lille. Y.Yuan acknowledges the financial support from the Henan Provincial Hospital., ANR-13-BSV8-0002,MECA VENUS,Mécanismes moléculaires de la transduction de signal par BvgS, un modèle de la famille de récepteurs kinases bactériens à domaines Vénus Flytrap(2013), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), and Jacob-Dubuisson, Françoise
- Subjects
Bordetella pertussis ,sensor kinases ,[SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,Mutagenesis (molecular biology technique) ,Biology ,Microbiology ,03 medical and health sciences ,PAS domain ,Molecular Biology ,030304 developmental biology ,Coiled coil ,0303 health sciences ,[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,030306 microbiology ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,Periplasmic space ,biology.organism_classification ,[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,Cell biology ,Regulon ,two-component system ,Signal transduction ,virulence regulation ,Function (biology) ,Research Article - Abstract
The two-component system BvgAS controls the virulence regulon in Bordetella pertussis. BvgS is the prototype of a family of sensor histidine kinases harboring periplasmic Venus flytrap (VFT) domains. The VFT domains are connected to the cytoplasmic kinase moiety by helical linkers separated by a Per-ARNT-Sim (PAS) domain. Antagonism between the two linkers, as one forms a coiled coil when the other is dynamic and vice versa, regulates BvgS activity. Here, we solved the structure of the intervening PAS domain by X-ray crystallography. Two forms were obtained that notably differ by the connections between the PAS core domain and the flanking helical linkers. Structure-guided mutagenesis indicated that those connections participate in the regulation of BvgS activity. Thus, the PAS domain appears to function as a switch facilitator module whose conformation determines the output of the system. As many BvgS homologs have similar architectures, the mechanisms unveiled here are likely to generally apply to the regulation of sensor histidine kinases of that family. IMPORTANCE The whooping cough agent Bordetella pertussis colonizes the human respiratory tract by using virulence factors coregulated by the sensory transduction system BvgAS. BvgS is a model for a family of sensor kinase proteins, some of which are found in important bacterial pathogens. BvgS functions as a kinase or a phosphatase depending on external signals, which determines if B. pertussis is virulent or avirulent. Thus, deciphering its mode of action might lead to new ways of fighting infections. Here, we used X-ray crystallography to solve the three-dimensional structure of the domain that precedes the enzymatic moiety and identified features that regulate BvgS activity. As many sensor kinases of the BvgS family harbor homologous domains, the mechanism unveiled here might be of general relevance.
- Published
- 2021
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