A chemical and biological toolbox for Type Vd secretion: Characterization of the phospholipase A1 autotransporter FplA from Fusobacterium nucleatum

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dc.contributor.authorCasasanta, Michael A.en
dc.contributor.authorYoo, Christopher C.en
dc.contributor.authorSmith, Hans B.en
dc.contributor.authorDuncan, Alison J.en
dc.contributor.authorCochrane, Kylaen
dc.contributor.authorVarano, Ann C.en
dc.contributor.authorAllen-Vercoe, Emmaen
dc.contributor.authorSlade, Daniel J.en
dc.contributor.departmentBiochemistryen
dc.contributor.departmentFralin Biomedical Research Instituteen
dc.date.accessioned2018-01-15T20:28:40Zen
dc.date.available2018-01-15T20:28:40Zen
dc.date.issued2017-12-08en
dc.description.abstractFusobacterium nucleatum is an oral pathogen that is linked to multiple human infections and colorectal cancer. Strikingly, F. nucleatum achieves virulence in the absence of large, multiprotein secretion systems (Types I, II, III, IV, and VI), which are widely used by Gram-negative bacteria for pathogenesis. By contrast, F. nucleatum strains contain genomic expansions of Type V secreted effectors (autotransporters) that are critical for host cell adherence, invasion, and biofilm formation. Here, we present the first characterization of an F. nucleatum Type Vd phospholipase class A1 autotransporter (strain ATCC 25586, gene FN1704) that we hereby rename Fusobacterium phospholipase autotransporter (FplA). Biochemical analysis of multiple Fusobacterium strains revealed that FplA is expressed as a fulllength 85-kDa outer membrane–embedded protein or as a truncated phospholipase domain that remains associated with the outer membrane. Whereas the role of Type Vd secretion in bacteria remains unidentified, we show that FplA binds with high affinity to host phosphoinositide-signaling lipids, revealing a potential role for this enzyme in establishing an F. nucleatum intracellular niche. To further analyze the role of FplA, we developed an fplA gene knock-out strain, which will guide future in vivo studies to determine its potential role in F. nucleatum pathogenesis. In summary, using recombinant FplA constructs, we have identified a biochemical toolbox that includes lipid substrates for enzymatic assays, potent inhibitors, and chemical probes to detect, track, and characterize the role of Type Vd secreted phospholipases in Gramnegative bacteria.en
dc.description.versionPublished versionen
dc.format.extent20240 - 20254 (15) page(s)en
dc.identifier.doihttps://doi.org/10.1074/jbc.M117.819144en
dc.identifier.eissn1083-351Xen
dc.identifier.issn0021-9258en
dc.identifier.issue49en
dc.identifier.orcidSlade, DJ [0000-0001-5634-7220]en
dc.identifier.urihttp://hdl.handle.net/10919/81789en
dc.identifier.volume292en
dc.language.isoenen
dc.publisherAmer Soc Biochemistry Molecular Biology Incen
dc.relation.urihttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000417696400028&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1en
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectBiochemistry & Molecular Biologyen
dc.subjectchemical biologyen
dc.subjectcolorectal canceren
dc.subjecthost-pathogen interactionen
dc.subjectphospholipase Aen
dc.subjectprotein secretionen
dc.subjectFusobacteriumen
dc.subjectautotransporteren
dc.subjectPSEUDOMONAS-AERUGINOSAen
dc.subjectINFECTIVE ENDOCARDITISen
dc.subjectSERINE HYDROLASESen
dc.subjectBRAIN ABSCESSESen
dc.subjectPRETERM BIRTHen
dc.subjectHOST-CELLSen
dc.subjectFAD-Ien
dc.subjectPROTEINen
dc.subjectINHIBITIONen
dc.subjectVIRULENCEen
dc.titleA chemical and biological toolbox for Type Vd secretion: Characterization of the phospholipase A1 autotransporter FplA from Fusobacterium nucleatumen
dc.title.serialJournal of Biological Chemistryen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciencesen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/Biochemistryen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/CALS T&R Facultyen
pubs.organisational-group/Virginia Tech/All T&R Facultyen
pubs.organisational-group/Virginia Tech/Faculty of Health Sciencesen

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