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  • Journal article
    Aurass P, Schlegel M, Metwally O, Harding CR, Schroeder GN, Frankel G, Flieger Aet al., 2013,

    The <i>Legionella pneumophila</i> Dot/Icm-secreted Effector PlcC/CegC1 Together with PlcA and PlcB Promotes Virulence and Belongs to a Novel Zinc Metallophospholipase C Family Present in Bacteria and Fungi

    , JOURNAL OF BIOLOGICAL CHEMISTRY, Vol: 288, Pages: 11080-11092
  • Journal article
    Figueira R, Watson KG, Holden DW, Helaine Set al., 2013,

    Identification of salmonella pathogenicity island-2 type III secretion system effectors involved in intramacrophage replication of S. enterica serovar typhimurium: implications for rational vaccine design

    , mBio, Vol: 4, ISSN: 2161-2129

    Salmonella enterica serovars cause severe diseases in humans, such as gastroenteritis and typhoid fever. The development of systemic disease is dependent on a type III secretion system (T3SS) encoded by Salmonella pathogenicity island-2 (SPI-2). Translocation of effector proteins across the Salmonella-containing vacuole, via the SPI-2 T3SS, enables bacterial replication within host cells, including macrophages. Here, we investigated the contribution of these effectors to intramacrophage replication of Salmonella enterica serovar Typhimurium using Fluorescence Dilution, a dual-fluorescence tool which allows direct measurement of bacterial replication. Of 32 strains, each carrying single mutations in genes encoding effectors, 10 (lacking sifA, sseJ, sopD2, sseG, sseF, srfH, sseL, spvD, cigR, or steD) were attenuated in replication in mouse bone marrow-derived macrophages. The replication profiles of strains combining deletions in effector genes were also investigated: a strain lacking the genes sseG, sopD2, and srfH showed an increased replication defect compared to single-mutation strains and was very similar to SPI-2 T3SS-deficient bacteria with respect to its replication defect. This strain was substantially attenuated in virulence in vivo and yet retained intracellular vacuole integrity and a functional SPI-2 T3SS. Moreover, this strain was capable of SPI-2 T3SS-mediated delivery of a model antigen for major histocompatibility complex (MHC) class I-dependent T-cell activation. This work establishes a basis for the use of a poly-effector mutant strain as an attenuated vaccine carrier for delivery of heterologous antigens directly into the cytoplasm of host cells.IMPORTANCE Live attenuated strains of Salmonella enterica serotype Typhi have generated much interest in the search for improved vaccines against typhoid fever and as vaccine vectors for the delivery of heterologous antigens. A promising vaccine candidate is the ΔaroC ΔssaV S. Typhi strain, whic

  • Journal article
    Sheppard C, James E, Barton G, Matthews S, Severinov K, Wigneshweraraj Set al., 2013,

    A non-bacterial transcription factor inhibits bacterial transcription by a multipronged mechanism

    , RNA BIOLOGY, Vol: 10, Pages: 495-501, ISSN: 1547-6286
  • Journal article
    Helaine S, Holden DW, 2013,

    Heterogeneity of intracellular replication of bacterial pathogens

    , CURRENT OPINION IN MICROBIOLOGY, Vol: 16, Pages: 184-191, ISSN: 1369-5274
  • Journal article
    Permpoonpattana P, Phetcharaburanin J, Mikelsone A, Dembek M, Tan S, Brisson M-C, La Ragione R, Brisson AR, Fairweather N, Hong HA, Cutting SMet al., 2013,

    Functional Characterization of <i>Clostridium difficile</i> Spore Coat Proteins

    , JOURNAL OF BACTERIOLOGY, Vol: 195, Pages: 1492-1503, ISSN: 0021-9193
  • Journal article
    Hachani A, Lossi NS, Filloux A, 2013,

    A visual assay to monitor T6SS-mediated bacterial competition

    , Jove-Journal of Visualized Experiments, ISSN: 1940-087X

    Type VI secretion systems (T6SSs) are molecular nanomachines allowing Gram-negative bacteria to transport and inject proteins into a wide variety of target cells1,2. The T6SS is composed of 13 core components and displays structural similarities with the tail-tube of bacteriophages3. The phage uses a tube and a puncturing device to penetrate the cell envelope of target bacteria and inject DNA. It is proposed that the T6SS is an inverted bacteriophage device creating a specific path in the bacterial cell envelope to drive effectors and toxins to the surface. The process could be taken further and the T6SS device could perforate other cells with which the bacterium is in contact, thus injecting the effectors into these targets. The tail tube and puncturing device parts of the T6SS are made with Hcp and VgrG proteins, respectively4,5.The versatility of the T6SS has been demonstrated through studies using various bacterial pathogens. The Vibrio cholerae T6SS can remodel the cytoskeleton of eukaryotic host cells by injecting an "evolved" VgrG carrying a C-terminal actin cross-linking domain6,7. Another striking example was recently documented using Pseudomonas aeruginosa which is able to target and kill bacteria in a T6SS-dependent manner, therefore promoting the establishment of bacteria in specific microbial niches and competitive environment8,9,10.In the latter case, three T6SS-secreted proteins, namely Tse1, Tse2 and Tse3 have been identified as the toxins injected in the target bacteria (Figure 1). The donor cell is protected from the deleterious effect of these effectors via an anti-toxin mechanism, mediated by the Tsi1, Tsi2 and Tsi3 immunity proteins8,9,10. This antimicrobial activity can be monitored when T6SS-proficient bacteria are co-cultivated on solid surfaces in competition with other bacterial species or with T6SS-inactive bacteria of the same species8,11,12,13.The data available emphasized a numerical approach to the bacterial competition assay

  • Journal article
    Firdessa R, Berg S, Hailu E, Schelling E, Gumi B, Erenso G, Gadisa E, Kiros T, Habtamu M, Hussein J, Zinsstag J, Robertson BD, Ameni G, Lohan AJ, Loftus B, Comas I, Gagneux S, Tschopp R, Yamuah L, Hewinson G, Gordon SV, Young DB, Aseffa Aet al., 2013,

    Mycobacterial Lineages Causing Pulmonary and Extrapulmonary Tuberculosis, Ethiopia

    , Emerging Infectious Diseases, Vol: 19, Pages: 460-463, ISSN: 1080-6059

    Molecular typing of 964 specimens from patients in Ethiopia with lymph node or pulmonary tuberculosis showed a similar distribution of Mycobacterium tuberculosis strains between the 2 disease manifestations and a minimal role for M. bovis. We report a novel phylogenetic lineage of M. tuberculosis strongly associated with the Horn of Africa.

  • Journal article
    Clare S, John V, Walker AW, Hill JL, Abreu-Goodger C, Hale C, Goulding D, Lawley TD, Mastroeni P, Frankel G, Enright AJ, Vigorito E, Dougan Get al., 2013,

    Enhanced Susceptibility to <i>Citrobacter rodentium</i> Infection in MicroRNA-155-Deficient Mice

    , INFECTION AND IMMUNITY, Vol: 81, Pages: 723-732, ISSN: 0019-9567
  • Journal article
    Williams KJ, Bennett MH, Barton GR, Jenkins VA, Robertson BDet al., 2013,

    Adenylylation of mycobacterial Glnk (PII) protein is induced by nitrogen limitation

    , TUBERCULOSIS, Vol: 93, Pages: 198-206, ISSN: 1472-9792
  • Journal article
    Shadrin A, Sheppard C, Savalia D, Severinov K, Wigneshweraraj Set al., 2013,

    Overexpression of <i>Escherichia coli udk</i> mimics the absence of T7 Gp2 function and thereby abrogates successful infection by T7 phage

    , MICROBIOLOGY-SGM, Vol: 159, Pages: 269-274, ISSN: 1350-0872

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