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2009-05-01Zeitschriftenartikel DOI: 10.1128/AEM.00187-09
bdhA-patD operon as a virulence determinant, revealed by a novel large-scale approach for identification of Legionella pneumophila mutants defective for amoeba infection.
dc.contributor.authorAurass, Philipp
dc.contributor.authorPless, Birgit
dc.contributor.authorRydzewski, Kerstin
dc.contributor.authorHolland, Gudrun
dc.contributor.authorBannert, Norbert
dc.contributor.authorFlieger, Antje
dc.date.accessioned2018-05-07T14:29:13Z
dc.date.available2018-05-07T14:29:13Z
dc.date.created2011-03-15
dc.date.issued2009-05-01none
dc.identifier.otherhttp://edoc.rki.de/oa/articles/reriWUD6hZmto/PDF/25oLgqSmVQSNw.pdf
dc.identifier.urihttp://edoc.rki.de/176904/836
dc.description.abstractLegionella pneumophila, the causative agent of Legionnaires' disease, is an intracellular parasite of eukaryotic cells. In the environment, it colonizes amoebae. After being inhaled into the human lung, the bacteria infect and damage alveolar cells in a way that is mechanistically similar to the amoeba infection. Several L. pneumophila traits, among those the Dot/Icm type IVB protein secretion machinery, are essential for exploiting host cells. In our search for novel Legionella virulence factors, we developed an agar plate assay, designated the scatter screen, which allowed screening for mutants deficient in infecting Acanthamoeba castellanii amoebae. Likewise, an L. pneumophila clone bank consisting of 23,000 transposon mutants was investigated here, and 19 different established Legionella virulence genes, for example, dot/icm genes, were identified. Importantly, 70 novel virulence-associated genes were found. One of those is L. pneumophila bdhA, coding for a protein with homology to established 3-hydroxybutyrate dehydrogenases involved in poly-3-hydroxybutyrate metabolism. Our study revealed that bdhA is cotranscribed with patD, encoding a patatin-like protein of L. pneumophila showing phospholipase A and lysophospholipase A activities. In addition to strongly reduced lipolytic activities and increased poly-3-hydroxybutyrate levels, the L. pneumophila bdhA-patD mutant showed a severe replication defect in amoebae and U937 macrophages. Our data suggest that the operon is involved in poly-3-hydroxybutyrate utilization and phospholipolysis and show that the bdhA-patD operon is a virulence determinant of L. pneumophila. In summary, the screen for amoeba-sensitive Legionella clones efficiently isolated mutants that do not grow in amoebae and, in the case of the bdhA-patD mutant, also human cells.eng
dc.language.isoeng
dc.publisherRobert Koch-Institut, Infektionskrankheiten / Erreger
dc.subjectCell Lineeng
dc.subjectHumanseng
dc.subjectVirulenceeng
dc.subjectAnimalseng
dc.subjectBacterial Proteins/geneticseng
dc.subjectLegionella pneumophila/pathogenicityeng
dc.subjectDNA Transposable Elementseng
dc.subjectAcanthamoeba castellanii/microbiologyeng
dc.subjectHydroxybutyrate Dehydrogenase/geneticseng
dc.subjectMacrophages/microbiologyeng
dc.subjectMutagenesis Insertionaleng
dc.subjectOperoneng
dc.subjectPhospholipases/geneticseng
dc.subjectVirulence Factors/geneticseng
dc.subject.ddc610 Medizin
dc.titlebdhA-patD operon as a virulence determinant, revealed by a novel large-scale approach for identification of Legionella pneumophila mutants defective for amoeba infection.
dc.typeperiodicalPart
dc.identifier.urnurn:nbn:de:0257-10012857
dc.identifier.doi10.1128/AEM.00187-09
dc.identifier.doihttp://dx.doi.org/10.25646/761
local.edoc.container-titleApplied and Environmental Microbiology
local.edoc.container-textAurass, P., Pless, B., Rydzewski, K., Holland, G., Bannert, N., Flieger, A. bdhA-patD operon as a virulence determinant, revealed by a novel large-scale approach for identification of Legionella pneumophila mutants defective for amoeba infection (2009) Applied and Environmental Microbiology, 75 (13), pp. 4506-4515.
local.edoc.fp-subtypeArtikel
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttp://aem.asm.org/cgi/content/full/75/13/4506?view=long&pmid=19411431
local.edoc.container-publisher-nameAmerican Society for Microbiology
local.edoc.container-volume75
local.edoc.container-issue13
local.edoc.container-year2009

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