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

Persistent Link:
http://hdl.handle.net/10150/610055
Title:
Phospholipase D promotes Arcanobacterium haemolyticum adhesion via lipid raft remodeling and host cell death following bacterial invasion
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
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

Full metadata record

DC FieldValue Language
dc.contributor.authorLucas, Erynnen
dc.contributor.authorBillington, Stephenen
dc.contributor.authorCarlson, Petterien
dc.contributor.authorMcGee, Daviden
dc.contributor.authorJost, B. H.en
dc.date.accessioned2016-05-20T08:57:32Z-
dc.date.available2016-05-20T08:57:32Z-
dc.date.issued2010en
dc.identifier.citationLucas et al. BMC Microbiology 2010, 10:270 http://www.biomedcentral.com/1471-2180/10/270en
dc.identifier.doi10.1186/1471-2180-10-270en
dc.identifier.urihttp://hdl.handle.net/10150/610055-
dc.description.abstractBACKGROUND: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.en
dc.language.isoenen
dc.publisherBioMed Centralen
dc.relation.urlhttp://www.biomedcentral.com/1471-2180/10/270en
dc.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)en
dc.titlePhospholipase D promotes Arcanobacterium haemolyticum adhesion via lipid raft remodeling and host cell death following bacterial invasionen
dc.typeArticleen
dc.identifier.eissn1471-2180en
dc.contributor.departmentDepartment of Veterinary Science and Microbiology, The University of Arizona, 1117 E Lowell Street, Tucson, AZ 85721, USAen
dc.contributor.departmentDepartment of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Haartmaninkatu 3, P.O. Box 21, Helsinki, FIN-00014, Finlanden
dc.contributor.departmentDepartment of Microbiology and Immunology, Louisiana State University Health Sciences Center-Shreveport, 1501 Kings Highway, Shreveport, LA 71130, USAen
dc.contributor.departmentModesto Junior College, 435 College Avenue, Modesto, CA 95350, USAen
dc.contributor.departmentVentana Medical Systems, Inc., 1910 Innovation Park Drive, Oro Valley, AZ 85755, USAen
dc.contributor.departmentOffice for the Responsible Conduct of Research, The University of Arizona, P.O. Box 245092, Tucson, AZ 85721, USAen
dc.identifier.journalBMC Microbiologyen
dc.description.collectioninformationThis 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.en
dc.eprint.versionFinal published versionen
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