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  • Journal article
    Muhl D, Filloux A, 2014,

    Site-Directed Mutagenesis and Gene Deletion Using Reverse Genetics

    , PSEUDOMONAS: METHODS AND PROTOCOLS, Vol: 1149, Pages: 521-539, ISSN: 1064-3745
  • Journal article
    Filloux A, Ramos J-L, 2014,

    Pseudomonas Methods and Protocols Preface

    , PSEUDOMONAS: METHODS AND PROTOCOLS, Vol: 1149, Pages: V-V, ISSN: 1064-3745
  • Journal article
    Filloux A, 2013,

    Fit and resistant is a worst case scenario with bacterial pathogens

    , PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol: 110, Pages: 20360-20361, ISSN: 0027-8424
  • Journal article
    Jones C, Hachani A, Manoli E, Filloux Aet al., 2013,

    An rhs Gene Linked to the Second Type VI Secretion Cluster Is a Feature of the Pseudomonas aeruginosa Strain PA14

    , Journal of Bacteriology, Vol: 196, Pages: 800-810, ISSN: 1098-5530

    The type VI secretion system (T6SS) of Gram-negative bacteria has been involved in various processes, notably bacterial competition and eukaryotic cell subversion. Most Pseudomonas aeruginosa strains possess three T6SS gene clusters, but only the function of the first T6SS (H1-T6SS) has been clearly elucidated. It is involved in the secretion of three toxins (Tse1 to -3) that target bacterial competitors. In the case of the H2- and H3-T6SS, no clear function has been assigned, and only one effector has been associated with these systems. Yet the H2-T6SS was proposed to promote P. aeruginosa internalization in nonphagocytic epithelial cells. Although the H2-T6SS genetic organization is conserved across P. aeruginosa isolates, one feature is the presence of an additional transcriptional unit in the PA14 strain H2-T6SS cluster, which is divergent from the core H2-T6SS genes. A specific set of four genes encodes an Hcp protein (Hcp2), a VgrG protein (VgrG14), an Rhs element (PA14_43100 or RhsP2), and a protein with no homologies with previously characterized proteins (PA14_43090). In this study, we engineered a P. aeruginosa PA14 strain carrying an arabinose-inducible H2-T6SS on the chromosome. We showed that arabinose induction readily promotes assembly of the H2-T6SS, as seen by monitoring Hcp2 secretion. We further studied the secretion fate of VgrG14 and RhsP2, but these were not detectable in the extracellular medium. We finally investigated whether activation of the PA14 H2-T6SS gene cluster could influence phenotypic traits such as internalization in eukaryotic cells, and we reported noteworthy differences compared to strain PAO1, which may be accounted for by the described genetic differences.

  • Journal article
    Bae B, Davis E, Brown D, Campbell EA, Wigneshweraraj S, Darst SAet al., 2013,

    Phage T7 Gp2 inhibition of Escherichia coli RNA polymerase involves misappropriation of σ70 domain 1.1.

    , Proceedings of the National Academy of Sciences, Vol: 110, Pages: 19772-19777
  • Journal article
    James EH, Edwards AM, Wigneshweraraj S, 2013,

    Transcriptional downregulation of agr expression in Staphylococcus aureus during growth in human serum can be overcome by constitutively active mutant forms of the sensor kinase AgrC

    , FEMS Microbiology Letters, Vol: 349, Pages: 153-162, ISSN: 0378-1097

    The temporal and cell density-dependent regulation of expression of virtually all the Staphylococcus aureus virulon is under the control of the agr (accessory gene regulatory) operon. The expression of the agr operon is subject to transcriptional regulation by the AgrA/C two-component response regulator/sensor kinase pair. During bacteraemia, a frequent syndrome caused by methicillin-resistant S. aureus (MRSA), the transcriptional downregulation of agr expression has been attributed to the sequestration of the quorum-signalling molecule auto-inducing peptide (AIP) by the human serum component apolipoprotein B as part of an innate immune response to infection. However, it is not known whether transcriptional downregulation of agr expression during growth in human serum is additionally subjected to regulation by transcription regulatory proteins that either directly or indirectly affect transcription from the agr operon promoters. Here, using chromosomal fluorescence reporters of agr expression in S. aureus, we show that the transcriptional downregulation of agr expression in human serum can be overcome using constitutive active mutant forms of AgrC. Therefore, it seems that the sequestration of the AIP is likely to be the only mechanism by which the host innate immune response limits agr expression at the transcriptional level to maintain the host–pathogen balance towards a noninvasive outcome.

  • Conference paper
    Al Shammari B, Shiomi T, Tezera L, Workman V, Jayasinghe S, Sathyamoorthy T, Mauri F, Robertson BD, Friedland JS, D'Armiento J, Elkington PTet al., 2013,

    Cell-matrix interactions regulate the immune response to Mycobacterium tuberculosis

    , Annual Congress of the British-Society-for-Immunology, Publisher: WILEY-BLACKWELL, Pages: 104-104, ISSN: 0019-2805
  • Journal article
    Muniz-Feliciano L, Van Grol J, Portillo J-AC, Liew L, Liu B, Carlin CR, Carruthers VB, Matthews S, Subauste CSet al., 2013,

    <i>Toxoplasma gondii</i>-Induced Activation of EGFR Prevents Autophagy Protein-Mediated Killing of the Parasite

    , PLOS PATHOGENS, Vol: 9, ISSN: 1553-7366
  • Journal article
    Schumacher J, Behrends V, Pan Z, Brown DR, Heydenreich F, Lewis MR, Bennett MH, Razzaghi B, Komorowski M, Barahona M, Stumpf MPH, Wigneshweraraj S, Bundy JG, Buck Met al., 2013,

    Nitrogen and Carbon Status Are Integrated at the Transcriptional Level by the Nitrogen Regulator NtrC <i>In Vivo</i>

    , MBIO, Vol: 4, ISSN: 2150-7511
  • Journal article
    Lu Z, Bergeron JRC, Atkinson RA, Schaller T, Veselkov DA, Oregioni A, Yang Y, Matthews SJ, Malim MH, Sanderson MRet al., 2013,

    Insight into the HIV-1 Vif SOCS-box-ElonginBC interaction

    , Open Biology, Vol: 3, Pages: 1-11, ISSN: 2046-2441

    The HIV-1 viral infectivity factor (Vif) neutralizes cell-encoded antiviral APOBEC3 proteins by recruiting a cellular ElonginB (EloB)/ElonginC (EloC)/Cullin5-containing ubiquitin ligase complex, resulting in APOBEC3 ubiquitination and proteolysis. The suppressors-of-cytokine-signalling-like domain (SOCS-box) of HIV-1 Vif is essential for E3 ligase engagement, and contains a BC box as well as an unusual proline-rich motif. Here, we report the NMR solution structure of the Vif SOCS–ElonginBC (EloBC) complex. In contrast to SOCS-boxes described in other proteins, the HIV-1 Vif SOCS-box contains only one α-helical domain followed by a β-sheet fold. The SOCS-box of Vif binds primarily to EloC by hydrophobic interactions. The functionally essential proline-rich motif mediates a direct but weak interaction with residues 101–104 of EloB, inducing a conformational change from an unstructured state to a structured state. The structure of the complex and biophysical studies provide detailed insight into the function of Vif's proline-rich motif and reveal novel dynamic information on the Vif–EloBC interaction.

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