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dc.contributor.authorHalbedel, Sven
dc.contributor.authorReiss, Swantje
dc.contributor.authorHahn, Birgit
dc.contributor.authorAlbrecht, Dirk
dc.contributor.authorMannala, Gopala Krishna
dc.contributor.authorChakraborty, Trinad
dc.contributor.authorHain, Torsten
dc.contributor.authorEngelmann, Susanne
dc.contributor.authorFlieger, Antje
dc.date.accessioned2015-01-23T13:11:06Z
dc.date.available2015-01-23T13:11:06Z
dc.date.issued2014-11
dc.identifier.citationA systematic proteomic analysis of Listeria monocytogenes house-keeping protein secretion systems. 2014, 13 (11):3063-81 Mol. Cell Proteomicsen
dc.identifier.issn1535-9484
dc.identifier.pmid25056936
dc.identifier.doi10.1074/mcp.M114.041327
dc.identifier.urihttp://hdl.handle.net/10033/338723
dc.description.abstractListeria monocytogenes is a firmicute bacterium causing serious infections in humans upon consumption of contaminated food. Most of its virulence factors are secretory proteins either released to the medium or attached to the bacterial surface. L. monocytogenes encodes at least six different protein secretion pathways. Although great efforts have been made in the past to predict secretory proteins and their secretion routes using bioinformatics, experimental evidence is lacking for most secretion systems. Therefore, we constructed mutants in the main housekeeping protein secretion systems, which are the Sec-dependent transport, the YidC membrane insertases SpoIIIJ and YqjG, as well as the twin-arginine pathway, and analyzed their secretion and virulence defects. Our results demonstrate that Sec-dependent secretion and membrane insertion of proteins via YidC proteins are essential for viability of L. monocytogenes. Depletion of SecA or YidC activity severely affected protein secretion, whereas loss of the Tat-pathway was without any effect on secretion, viability, and virulence. Two-dimensional gel electrophoresis combined with protein identification by mass spectrometry revealed that secretion of many virulence factors and of enzymes synthesizing and degrading the cell wall depends on the SecA route. This finding was confirmed by SecA inhibition experiments using sodium azide. Analysis of secretion of substrates typically dependent on the accessory SecA2 ATPase in wild type and azide resistant mutants of L. monocytogenes revealed for the first time that SecA2-dependent protein secretion also requires the ATPase activity of the house-keeping SecA protein.
dc.language.isoenen
dc.titleA systematic proteomic analysis of Listeria monocytogenes house-keeping protein secretion systems.en
dc.typeArticleen
dc.identifier.journalMolecular & cellular proteomics : MCPen
refterms.dateFOA2015-11-15T00:00:00Z
html.description.abstractListeria monocytogenes is a firmicute bacterium causing serious infections in humans upon consumption of contaminated food. Most of its virulence factors are secretory proteins either released to the medium or attached to the bacterial surface. L. monocytogenes encodes at least six different protein secretion pathways. Although great efforts have been made in the past to predict secretory proteins and their secretion routes using bioinformatics, experimental evidence is lacking for most secretion systems. Therefore, we constructed mutants in the main housekeeping protein secretion systems, which are the Sec-dependent transport, the YidC membrane insertases SpoIIIJ and YqjG, as well as the twin-arginine pathway, and analyzed their secretion and virulence defects. Our results demonstrate that Sec-dependent secretion and membrane insertion of proteins via YidC proteins are essential for viability of L. monocytogenes. Depletion of SecA or YidC activity severely affected protein secretion, whereas loss of the Tat-pathway was without any effect on secretion, viability, and virulence. Two-dimensional gel electrophoresis combined with protein identification by mass spectrometry revealed that secretion of many virulence factors and of enzymes synthesizing and degrading the cell wall depends on the SecA route. This finding was confirmed by SecA inhibition experiments using sodium azide. Analysis of secretion of substrates typically dependent on the accessory SecA2 ATPase in wild type and azide resistant mutants of L. monocytogenes revealed for the first time that SecA2-dependent protein secretion also requires the ATPase activity of the house-keeping SecA protein.


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