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    Phospholipase D promotes Arcanobacterium haemolyticum adhesion via lipid raft remodeling and host cell death following bacterial invasion

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    Author
    Lucas, Erynn
    Billington, Stephen
    Carlson, Petteri
    McGee, David
    Jost, B. H.
    Affiliation
    Department of Veterinary Science and Microbiology, The University of Arizona, 1117 E Lowell Street, Tucson, AZ 85721, USA
    Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Haartmaninkatu 3, P.O. Box 21, Helsinki, FIN-00014, Finland
    Department of Microbiology and Immunology, Louisiana State University Health Sciences Center-Shreveport, 1501 Kings Highway, Shreveport, LA 71130, USA
    Modesto Junior College, 435 College Avenue, Modesto, CA 95350, USA
    Ventana Medical Systems, Inc., 1910 Innovation Park Drive, Oro Valley, AZ 85755, USA
    Office for the Responsible Conduct of Research, The University of Arizona, P.O. Box 245092, Tucson, AZ 85721, USA
    Issue Date
    2010
    
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    Show full item record
    Publisher
    BioMed Central
    Citation
    Lucas et al. BMC Microbiology 2010, 10:270 http://www.biomedcentral.com/1471-2180/10/270
    Journal
    BMC Microbiology
    Rights
    © 2010 Lucas et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0)
    Collection Information
    This item is part of the UA Faculty Publications collection. For more information this item or other items in the UA Campus Repository, contact the University of Arizona Libraries at repository@u.library.arizona.edu.
    Abstract
    BACKGROUND:Arcanobacterium haemolyticum is an emerging bacterial pathogen, causing pharyngitis and more invasive infections. This organism expresses an unusual phospholipase D (PLD), which we propose promotes bacterial pathogenesis through its action on host cell membranes. The pld gene is found on a genomic region of reduced %G + C, suggesting recent horizontal acquisition.RESULTS:Recombinant PLD rearranged HeLa cell lipid rafts in a dose-dependent manner and this was inhibited by cholesterol sequestration. PLD also promoted host cell adhesion, as a pld mutant had a 60.3% reduction in its ability to adhere to HeLa cells as compared to the wild type. Conversely, the pld mutant appeared to invade HeLa cells approximately two-fold more efficiently as the wild type. This finding was attributable to a significant loss of host cell viability following secretion of PLD from intracellular bacteria. As determined by viability assay, only 15.6% and 82.3% of HeLa cells remained viable following invasion by the wild type or pld mutant, respectively, as compared to untreated HeLa cells. Transmission electron microscopy of HeLa cells inoculated with A. haemolyticum strains revealed that the pld mutant was contained within intracellular vacuoles, as compared to the wild type, which escaped the vacuole. Wild type-infected HeLa cells also displayed the hallmarks of necrosis. Similarly inoculated HeLa cells displayed no signs of apoptosis, as measured by induction of caspase 3/7, 8 or 9 activities.CONCLUSIONS:These data indicate that PLD enhances bacterial adhesion and promotes host cell necrosis following invasion, and therefore, may be important in the disease pathogenesis of A. haemolyticum infections.
    EISSN
    1471-2180
    DOI
    10.1186/1471-2180-10-270
    Version
    Final published version
    Additional Links
    http://www.biomedcentral.com/1471-2180/10/270
    ae974a485f413a2113503eed53cd6c53
    10.1186/1471-2180-10-270
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