Type III Secretion Effectors with Arginine N-Glycosyltransferase Activity
Abstract
Type III secretion systems are used by many Gram-negative bacterial pathogens to inject proteins, known as effectors, into the cytosol of host cells. These virulence factors interfere with a diverse array of host signal transduction pathways and cellular processes. Many effectors have catalytic activities to promote post-translational modifications of host proteins. This review focuses on a family of effectors with glycosyltransferase activity that catalyze addition of N-acetyl-d-glucosamine to specific arginine residues in target proteins, leading to reduced NF-κB pathway activation and impaired host cell death. This family includes NleB from Citrobacter rodentium, NleB1 and NleB2 from enteropathogenic and enterohemorrhagic Escherichia coli, and SseK1, SseK2, and SseK3 from Salmonella enterica. First, we place these effectors in the general framework of the glycosyltransferase superfamily and in the particular context of the role of glycosylation in bacterial pathogenesis. Then, we provide detailed information about currently known members of this family, their role in virulence, and their targets
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Microorganisms2020,8,357;doi:10.3390/microorganisms8030357www.mdpi.com/journal/microorganisms Review TypeIIISecretionEffectorswithArginine N‐GlycosyltransferaseActivity JuanLuisAraujo‐Garrido,JoaquínBernal‐BayardandFranciscoRamos‐Morales* DepartamentodeGenética,FacultaddeBiología,UniversidaddeSevilla,41012Sevilla,Spain; [email protected](J.L.A.‐G.)[email protected](J.B.‐B.) *Correspondence:[email protected] Received:12February2020;Accepted:29February2020;Published:2March2020 Abstract:TypeIIIsecretionsystemsareusedbymanyGram‐negativebacterialpathogenstoinject proteins,knownaseffectors,intothecytosolofhostcells.Thesevirulencefactorsinterferewitha diversearrayofhostsignaltransductionpathwaysandcellularprocesses.Manyeffectorshave catalyticactivitiestopromotepost‐translationalmodificationsofhostproteins.Thisreviewfocuses onafamilyofeffectorswithglycosyltransferaseactivitythatcatalyzeadditionofN‐acetyl‐ D ‐ glucosaminetospecificarginineresiduesintargetproteins,leadingtoreducedNF‐κBpathway activationandimpairedhostcelldeath.ThisfamilyincludesNleBfromCitrobacterrodentium,NleB1 andNleB2fromenteropathogenicandenterohemorrhagicEscherichiacoli,andSseK1,SseK2,and SseK3fromSalmonellaenterica.First,weplacetheseeffectorsinthegeneralframeworkofthe glycosyltransferasesuperfamilyandintheparticularcontextoftheroleofglycosylationinbacterial pathogenesis.Then,weprovidedetailedinformationaboutcurrentlyknownmembersofthis family,theirroleinvirulence,andtheirtargets. Keywords:glycosyltransferases;typeIIIsecretion;effectors;NleB;SseK;Citrobacter;Escherichia; Salmonella;deathdomains 1.Introduction TypeIIIsecretionsystems(T3SSs)arefundamentaltoolsfortheinteractionbetweenmany pathogenicandsymbioticGram‐negativebacteriaandtheirhosts.T3SSs,alsoknownasinjectisomes, spanthebacterialinnerandoutermembranesandforminjectionnanomachinesthatdeliverproteins, knownaseffectors,intotargeteukaryoticcellstomodulateavarietyofcellularfunctions[1,2]. Effectorshavediverseactivitiesandtargets,andtypicallyhaveanN‐terminalsecretiondomainand oneormoreC‐terminalfunctionaldomains[3,4].Effectorscanbeclassifiedinfournon‐mutually exclusivecategoriesaccordingtotheirmechanismsofactivity[5]:(i)effectorsthatmediatetheir activitiesthroughphysicalinteractionswithhosttargets;(ii)effectorsthatusefunctionalorstructural mimicrytoalterhostcellprocesses;(iii)effectorsthatpromotepost‐translationalmodificationsof hostproteins;and(iv)effectorswithproteaseactivity.Examplesofcatalyticactivitiesofeffectorsare ubiquitination,deubiquitination,phosphorylation,dephosphorylation,acetylation,deamidation, ADPribosylation,cysteinemethylation,andglycosylation.Thisreviewfocusesonaspecificfamily ofT3SSeffectorswithglycosyltransferaseactivitythatarefoundinSalmonellaentericaandin attaching/effacing(A/E)pathogens. SalmonellaearefacultativeintracellularGram‐negativebacterialpathogensbelongingtothe familyEnterobacteriaceae.ThegenusSalmonellaincludesthespeciesS.bongoriandS.enterica,the latterdividedintosixsubspeciesandmorethan2500serovars[6].S.entericapossessestwovirulencerelatedT3SS[7–9],T3SS1andT3SS2,thatareencodedbySalmonellapathogenicityisland1(SPI1)and Salmonellapathogenicityisland2(SPI2)(Figure1),respectively,whereasS.bongorilacksSPI2[10].
Microorganisms2020,8,3572of23 TheT3SS1translocateseffectorsacrossthehostcellplasmamembranethatpromoteSalmonella invasionintoepithelialcellsintheintestineandstimulatelocalizedinflammation[11].TheT3SS2is expressedintracellularlyinresponsetoacidicpHandnutrientlimitationfoundinthelumenofthe Salmonella‐containingvacuole(SCV),amodifiedphagolysosomewhichisthetypicalintracellular nicheofthispathogen[12].T3SS2effectorsaretranslocatedthroughtheSCVmembrane,are importantinintestinalanddisseminatedinfections,andarerequiredforgrowthwithindifferenthost celltypes[13].Together,T3SS1andT3SS2secretemorethan40effectors,someencodedbySPI1or SPI2,butmostencodedoutsidetheseislands,inotherhorizontallyacquiredregionsofthegenome [14–17]. EnterohemorrhagicEscherichiacoli(EHEC)andenteropathogenicE.coli(EPEC)areextracellular humanpathogensthatattachtotheintestinalepitheliumandeffacebrushbordermicrovilliforming A/Elesions[18].EHECcauseshemorrhagiccolitisandhemolyticuremicsyndrome,whereasEPEC isacauseofinfantilediarrhea.CitrobacterrodentiumisanotherA/Epathogenwhichisthecausative agentofmurinetransmissiblecolonichyperplasia[19].LesionsinducedbyC.rodentiuminmiceare similartothosecausedbyEHECorEPECinhumansandthesepathogensshareanumberof virulencedeterminantsincludingthelocusofenterocyteeffacement(LEE)[20,21](Figure1).TheLEE isa34kbpathogenicityislandwith41genesthatencodeallthestructuralcomponentsofaT3SS, sevenofitseffectorsandtheirchaperones,proteinsinvolvedinadherence,andregulatoryproteins [22].ManyeffectorsareencodedoutsidetheLEE,andthetotalnumberofeffectorsisatleast29inC. rodentium,22inEPEC,and39inEHEC[22].Asecondgenecluster,potentiallyencodingaT3SSand knownasETT2,forE.coliT3SS2,ispresentinmanyE.colistrains[23,24].ETT2resemblesthe SalmonellaT3SS1butinmostcasesithasbeensubjectedtomutationalattritionandthereisno evidenceforthesecretionofeffectorsbythissystem[25].However,thisregionisintactinsome strainsofE.coliandEscherichiaalbertii,andappearstobeimportantforbacterialpathogenesis[25– 27]. Figure1.GeneticstructureofSalmonellapathogenicityisland1(SPI1)andSalmonellapathogenicity island2(SPI2)ofS.entericaserovarTyphimuriumstrain14028,andthelocusofenterocyteeffacement (LEE)islandofE.coliO127:H6strainE2348/69.Genesarecoloredaccordingtotheirfunctional categories[22,28].
Microorganisms2020,8,3573of23 2.Glycosyltransferases Glycosyltransferases(GTs)catalyzetheformationofglycosidebondsusinganactivateddonor sugarsubstratethatcontainsanucleosidephosphateoralipidphosphateleavinggroup[29]. Enzymesthatusesugarmono‐ordiphosphonucleotidesasdonorsareknownasLeloirGTs[30]. Non‐LeloirGTsusenon‐nucleotidedonors,suchaspolyprenolpyrophosphates,polyprenol phosphates,sugar‐1‐phosphates,orsugar‐1‐pyrophosphates.Sugarscanbetransferredtoavariety ofbiomoleculesincludingglycans,lipids,proteins,peptides,andsmallmolecules.Theresulting glycosidiclinkagecanbeanO‐,S‐,N‐orC‐covalentbondconnectingamonosaccharidetoanother residue. 2.1.Classification Traditionally,GTswereclassifiedonthebasisoftheirdonor,acceptor,andproductspecificity, accordingtotherecommendationsoftheInternationalUnionofBiochemistryandMolecularBiology (IUBMB),undertheEnzymeCommissionnumberEC.2.4.Giventhelimitationsofthissystem,anew schemethatinvolvestheclassificationofGTsintofamiliesonthebasisofaminoacidsequence similaritiesisnowaccepted[31,32].Today,thecarbohydrateactiveenzymes(CAZy)database (http://www.cazy.org)includesover620,000GTmodulesclassifiedinmorethan100familiesand morethan13,000nonclassifiedmodules. Dependingontheirmechanismoftransfer,GTscanbeclassifiedasinvertingorretaining, resultingininversion(α‐>β)orretention(α‐>α)ofthestereochemistryofthedonor’sanomeric bondduringthetransfer,respectively[33].TheoreticalandexperimentalstudiessupportasingledisplacementSN2(substitution,nucleophilic,bimolecular)‐likemechanismforinvertingGTs[29]. Thismechanisminvolvestheformationofanoxocarbeniumiontransitionstate(Figure2).Incontrast, thereactionmechanismofretainingglycosylationiscontroversial[34].Adouble‐displacement mechanism,inwhichthesugarisfirsttransferredtotheenzymeandthentransferredtotheacceptor, wassuggestedbyanalogytoretainingglycosidases[35].Someexperimentalsupportwasprovided forthismechanism[36,37],particularlyfortheGT6familymembersthathaveacarboxylateproperly orientedtoactasacatalyticnucleophile.ButthisisnotthecaseformostretainingGTs,leadingtothe proposalofanalternativeinternalreturnSNi(substitution,nucleophilic,internal)‐likemechanism, alsotermedfront‐facemechanism,inwhichthenucleophilichydroxygroupoftheacceptorattacks theanomericcarbonatomfromthesamesidefromwhichtheleavinggroupdeparts[38,39].An orthogonalmechanismhasalsobeenproposedthatdiffersfromtheSNimechanisminthereaction profileandthetimeofbondformationandbreakage[40](Figure2). Figure2.Glycosyltransferasemechanisms.AnSN2(substitution,nucleophilic,bimolecular)processis acceptedforinvertingglycosyltransferases(GTs).Severalmechanismshavebeenproposedfor retainingenzymes.Theorthogonalmechanismisdepictedhere.‡:Transitionstate.ImagebyDr.
Microorganisms2020,8,3574of23 BrockSchumann(WikimediaCommons,CCBY‐SA3.0,https://creativecommons.org/licenses/bysa/3.0/deed.en). 2.2.StructuralStudies GTstructures(Figure3)revealedthatthemajorityoftheseenzymesfallintotwogeneralfolds, calledGT‐AandGT‐B,eachofwhichincludesmembersthatareretainingandinvertingGTs.This ledtoaclassificationofGTsinfourclans:clanIforinvertingenzymeswiththeGT‐Afold,clanIIfor invertingenzymeswiththeGT‐Bfold,clanIIIforretainingenzymeswiththeGT‐Afold,andclanIV forretainingenzymeswiththeGT‐Bfold[32].BothfoldtypescontainaRossmannfold[41],a nucleotide‐bindingdomainresponsiblefordonornucleotiderecognition.Aseconddomainisusually responsibleforacceptorrecognitionandismorevariable.EnzymeswiththeGT‐Btypefoldcontain twoRossmann‐likefoldsseparatedbyadeepwidecrevice[42].However,notallGTsfitintothese clans.Athirdfoldtype,calledGT‐Cfold,waspredictedusingcomputationalmethods[43–45],and thestructureofseveralGTswiththisfold,mostofthembelongingtofamilyGT66,havebeen described[46].TheseGTsusuallycontainanN‐terminaltransmembranedomainandaC‐terminal globulardomainwithGTactivity.Inaddition,thestructureoftheenzymeDUF1792revealedanovel folddesignatedasGT‐Dtype[47]anddefinedfamilyGT101.MembersoftheGT51familyalsohave adistinctstructuredefinedaslysozyme‐type[48,49],andtherecentlypublishedstructureofTagA,a memberoftheGT26family,suggeststhedefinitionofanovelfoldtermedGT‐E[50].Acommon featurefoundinmanymembersoftheGT‐AandGT‐CsuperfamiliesisaDxDmotifinvolvedin bindingtodivalentmetalionandcatalysis.However,theoccurrenceofthismotifisnotenoughto predictapotentialGTfunctionsincemanyGTsdonotpossessitandtheDxDmotifisfoundinmore thanahalfofalldescribedproteins[32].Finally,giventhemodularnatureofcarbohydrate‐active enzymes,anumberofGTscontainnoncatalyticdomainspotentiallyinvolvedinsubstrate recognitionorprotein–proteininteraction,andsomepolypeptidescontaintwoGTmodules belongingtothesameordifferentfamilies. Figure3.Glycosyltransferasefolds.(a)DiagramoftheGT‐AfoldproteinSpsAfromBacillussubtilis, belongingtofamilyGT2(PDBID:1QG8)[51];(b)diagramoftheGT‐Bfold‐typeproteinGtfBfrom Amycolaptosisorientalis,belongingtofamilyGT1(PDBID:1IIR)[52];(c)diagramoftheGT‐Cfold proteinPglBfromCampylobacterlari,belongingtofamilyGT66(PDBID:3RCE)[46].Drawingscreated withNGL[53]. Table1showsaclassificationofGTfamiliesbasedonreactionmechanismsandfoldtypes.
Microorganisms2020,8,3575of23 Table1.Classificationofglycosyltransferasefamilies. ReactionMechanism InvertingRetainingUnknown Foldtype GT‐A21 , 7,12,13,14,16,21,29,31,42,43, 82,84,109 6,8,15,24,27, 34,44,55,62, 64,78,81,88 GT‐B1,9,10,18,19,23,28,30,33,38,41, 52,63,65,68,70,80,90,104 3,4,5,20,35, 52,72,99,107 GT‐C22,39,50,53,57,58,59,66,83,85,87 Others26(GT‐E),51(lysozyme‐type), 101(GT‐D) Unknown 11,17,25,37,40,47,48,49,54,56, 61,67,73,74,75,76,92,94,97,98, 100,102,103,105,106,108 32,45,60,69, 71,77,79,89, 93,95,96 91,110 1NumberscorrespondtotheGTfamilynumbersassignedinthecarbohydrateactiveenzymes (CAZy)database. 3.GlycosylationinBacterialPathogenesis Asmentionedintheintroduction,effectorsfrompathogenicbacteriaoftenexhibitspecific enzymaticactivityagainsthostcelltargetsaspartofitsinfectionstrategy.Proteinglycosylation,the mostcommonpost‐translationalmodification,hasbeentraditionallythoughttoberestrictedto eukaryoticorganisms.However,inthelastyears,manyexamplesofbacterialvirulencefactorsand toxinsexhibitingGTactivityhavebeencharacterized[54].Bacterialglycosylationisbroadlylinked tobacterialpathogenesis[55].Itfulfillstwomainfunctionsduringinfection:firstly,protein glycosylationenablesbacterialadhesiontohostcellsand,secondly,leadstomodificationofspecific hosttargetswiththeconsequentmanipulationofhostcellularprocessesbythepathogentotheirown benefit. BacterialGTsperformtwotypesofproteinmodifications:N‐linkedglycosylationorO‐linked glycosylation[29].WhereasN‐GTsconjugateglycanstosidechainamidenitrogenofanasparagine inanAsn‐X‐Ser/Thrconsensussequence,O‐glycosylationhappensonthehydroxylgroupofaserine orthreonineresidue.BothN‐linkedandO‐linkedglycosylationpathwaysmaymodifymultiple proteins.Interestingly,thereareexamplesofpathogenicbacteria,likeC.rodentium,performingN‐ andO‐glycosylationthatarebothcriticallyrequiredforcolonizationofthegastrointestinaltractin infectedmice[54]. InthissectionweproviderepresentativeexamplesofwellcharacterizedbacterialGTs,attending totheirspecificbiologicalroleduringinfectionandthetypeofmodificationtheyperform. 3.1.GlycosylationofAdhesins Oneessentialsteptoinitiateinfectionisbacterialadhesiontothehostepithelium.Thisfirststep dependssometimesonglycosylationofbacterialadhesins.Someexamplesaredescribedbelow. Awell‐knownexampleisthehigh‐molecular‐weight(HMW)systemofHaemophilusinfluenza. HMW1/HMW2aretwoadhesinsthatareglycosylatedatthecytoplasmbyGTsHMW1Cand HMW2C,respectively[56,57].Then,theyaretransportedtothebacterialsurfaceandmediate adherencetorespiratoryepithelialcells[57].Glucoseandgalactoseareaddedtospecific asparagineresiduesintheeukaryotic‐likeAsn‐X‐Ser/Thrconsensus[58]. Pgl(fromProteinglycosylation)systeminCampylobacterjejuniisanotherexampleofa glycosylatedadhesinthathasbeenconsideredaprototypefortherelevanceofbacterial
Microorganisms2020,8,3576of23 glycosylationinbacterial–hostinteraction[55].ItisanN‐linkedproteinglycosylationsystem composedofseveralproteins,includingvariousGTsthatmodifyC.jejuniproteinsleadingto bacterial–hostcelladhesionandinvasionofhostepithelium[59].DisruptionofthePglpathway resultsinreductionofhumanepithelialcellsadhesionanddecreasestheabilitytocolonize mouseandchickeninvivo[59–61].ModificationcatalyzedbythePglsystemconsistsofthe additionofaheptasaccharidetoasparagineresidueswithintheeukaryotic‐likeAsn‐X‐Ser/Thr glycosylationconsensus.Theoligosaccharideisassembledonthecytoplasmicsideoftheinner membrane.Then,itistranslocatedbyanABC‐transporter(PglK)[62].Onceintheperiplasmic space,PglB,anoligo‐saccharyltransferase,transferstheheptasaccharidetoasparagineresidues oftargetligands[63,64]. InadditiontoN‐glycosylation‐relatedmechanismstoimprovebacterialadhesion,othersystems basedonO‐glycosylationhavebeencharacterized.Oneexampleistheemergingbacterial autotransporterheptosyltransferase(BAHT)familyfoundinseveralGram‐negativebacteria [65].AutotransporterswhichbelongtotheT5SSfamilyhaveamodularcompositionwithanN‐ terminalpassengerdomainandaC‐terminalβ‐barreldomainthatdrivesthepassengeracross theoutermembrane.ThepassengerdomainmaybeheptosylatedbyaBAHTmemberon numerousserineresidues.Thismodificationoccursbeforetranslocationoftheproteinacrossthe innermembraneandisrequiredforbacterialadhesiontothehost.Twowell‐characterized BAHTmemberareAAH(autotransporteradhesinheptosyltransferase),fromdiffuselyadhering E.coliisolate2787,andTibCfromenterotoxigenicE.colistrainH10407[65,66].Inadditionto pathogenicE.coli,BAHTmembershavebeenidentifiedinotherbacteria,includingCitrobacter, Shigella,Salmonella,andBurkholderia[65].Furtherstudiesarenecessarytofullycharacterizethese interestingbacterialenzymes. Post‐translationalmodificationofbacterialproteinstoenhancebacterialadhesiontothehostis notrestrictedtoGram‐negativebacteria.Awell‐knownexampleofmodifiedbacterialproteins inGram‐positivebacteriaaretheO‐glycosylatedserine‐richrepeatproteins(SRRPs)[67].These largeglycoproteins,uniqueinGram‐positivebacteria,arecrucialforbiofilmformationand, moreimportantly,hostcelladhesion[68].Thebest‐characterizedSRRPisFap1from Streptococcusparasanguinis.GlycosylationofFap1reliesontheactivityofatleastsixdifferent GTs,althoughthespecificfunctionandglycantransferredremainsunclear[69].Thisfamilyof glycoproteinsishighlyconservedamongstreptococcalandstaphylococcalspecies[67],thus, characterizationofSRRPsGTsandunderstandingoftheirbiologicalrolearekeytoidentifynew therapeuticdrugtargetsinordertotacklerelevantGram‐positivepathogens. 3.2.GlycosylationofFlagellins GlycosylationofflagellinisanimportantcomponentofnumerousflagellarsystemsinArchaea andBacteria,playinganimportantroleinassemblyandvirulence[70],sincethisorganelle,in additiontoconferringmotility,canplayaroleineverystepoftheinfectioncycle[71].FlagellaO‐ glycosylationhasbeendemonstratedinmanypolarflagellinsfromGram‐negative[72]andGrampositivebacteria[73–80],andalsoinalimitednumberoflateralflagellins[72,81,82].Bacteriacan glycosylateflagellinatavaryingnumberofacceptorsites,fromasinglesiteinBurkholderiaspp.or ListeriamonocytogenestomultiplesitesinC.jejuniorSelenomonassputigena[80,83–85].Moreover, whereassomebacteriauseasinglemonosaccharide,othersshowstrain‐to‐strainglycan heterogenetity[72].Selectedexamplesaredescribedbelow. InAeromonascaviaeSch3N,thepolarflagellins,FlaAandFlaB,areglycosylatedwith pseudoaminicacidglycansandthisglycosylationisrequiredforflagellarassembly.Genes involvedinglycosylationaremappedintheO‐antigenbiosyntheticcluster[86,87].InAeromonas hydrophilaAH‐3,lateralflagellinismodifiedwithaderivativeofpseudaminicacid,whereas polarflagellaareglycosylatedwithaheterogeneousglycan[88]. C.jejuniFlaAisglycosylatedatupto19sitesthatcontributetotheflagellumserospecificity.FlaB isalsoglycosylated,butthenumberofsitesremainstobeestablished.ThepredominantO‐ glycansarederivativesofpseudoaminicacidandlegionaminicacid.ThegenesencodingGTs
Microorganisms2020,8,3577of23 involvedinflagellinglycosylationarelocatedadjacenttotheflagellinstructuralgenes,flaAand flaB[85,89,90]. InHelicobacterpylori,boththeFlaAandFlaBstructuralproteinsareglycosylatedwith pseudomainicacidatsevenandtensites,respectively.Biosynthesisofpseudaminicacidhas beenextensivelystudiedinH.pyloriandC.jejuniandinvolvessixconsecutiveenzymaticsteps beforebeingtransferredontoflagellinsviaO‐linkedglycosylation[91,92]. TheGTtransferringpseudaminicacidontoflagellinhasbeenstudiedinMagnetospirillum magneticumAMB‐1[93].ThisGT,knownasMaf,displayssignificantsequenceidentitywith proteinsfromotherGram‐negativebacteria,includingMaf1fromA.caviaeandPseEfromC. jejuni,thathavebeenshowntotransferpseudaminicacidontoflagellinsintheseorganisms [94,95].ThestructureofM.magneticumMafdisplaysamodifiedGT‐Atopologyasfoundin CAZyfamiliesGT29andGT42.Infact,thecloseststructuralhomologueisC.jejuni sialyltransferaseCst‐II(PDB1RO7),belongingtofamilyGT42[93]. GlycosyltationofClostridiumdifficileflagellinisessentialformotility.Twotypesofmodification havebeendefinedastypeAandB.TypeAmodificationinvolvesanO‐linkedN‐ acetylglucosamine(GlcNAc)linkedtoamethylatedthreonineviaaphosphodiesterbond.Three GTsareinvolvedintypeBmodification:GT1andGT2areresponsibleforthesequentialaddition ofaGlcNAcandtworhamnoses,respectively,andGT3isassociatedwiththeincorporationof asulfonatedpeptidyl‐amidosugarmoiety[74]. 3.3.ModificationofHostCellTargetsbyBacterialGTs InadditiontobacterialGTsthatglycosylatetheirownproteins,wefindanotherinteresting groupofbacterialproteinsthatmodifyhostcellproteins,manipulatingthecellphysiologyto counteractthehostdefenseresponse.ThelargeclostridialtoxinsTcdA/TcdB,theLegionellacytotoxic glycosyltransferasefamily(Lgt),andN‐GTsfromtheNleB/SseKfamilyareincludedinthisgroup [54]. Largeclostridialglycosylatingtoxins:pathogenesisofClostridiumdependsontheactivityoftwo similartoxins,TcdAandTcdB,thatexhibitGTactivityintheirN‐terminaldomains[96,97].Both toxinsperformO‐glycosylationofsmallGTPasesoftheRhofamily,inhibitingtheregulatory functionsoftheseproteins[98].Thismodificationoccursonthesidechainhydroxylgroupofa threonineresidue,whichisspecificforeachtargetedRhoGTPase[99].Anotherexampleofthis familyoftoxinsrecentlyidentifiedisPaToxfromentomopathogenicPhotorhabdusasymbiotica [100]. LegionellapneumophilaLgtfamily:L.pneumophilaisthecausativeagentoflegionellosis,asevere lungdisease.ThisfacultativeintracellularpathogenusesaT4SStoinjectmorethan300effectors intothehostcell[101].Amongthem,L.pneumophilasecretesseveralGTs(Lgt1‐3)thatperform specificglycosylationofaserineresidue(Ser53)intheeukaryoticelongationfactor1A,alarge GTPaseandacomponentoftheelongationcomplexinproteinsynthesis.Thismodification resultsininhibitionofproteinsynthesisandsubsequentlycausesthedeathofinfectedcells [102,103].Interestingly,Lgt1sharehighstructuralhomologywiththeclostridialtoxinTcdB. BothGTsaremembersoftheclanIIIofretainingenzymeswiththeGT‐Afold[103–105]. NleB/SseKfamily:werefertothisfamilyofT3SSeffectorsinthenextsection. 4.GlycosyltransferasesoftheNleBFamily 4.1.MembersoftheFamily T3SSeffectorsthatcatalyzethetransferofGlcNActoargininesofhostproteinshavebeen studiedinC.rodentium,EHEC,EPEC,andS.enterica(Table2).
Microorganisms2020,8,3578of23 Table2.NleB/SseKglycosyltransferases. Organism. Reference StrainEffectorNameProteinSize (aa)GenomicContextReferences C.rodentiumICC168NleB329GenomicislandGI4[106,107] Enterohemorrhagic E.coli(EHEC) O157:H7 EDL933 NleB1329O‐island122/SpLE3 [106,108] NleB2326O‐island36/PhageCP‐ 933K/Sp3 EnteropathogenicE. coli(EPEC)O127:H6E2348/69 NleB1329IntegrativeelementIE6 [109,110] NleB2326PhagePP4 S.entericaserovar Typhimurium14028 SseK1336 [111–113] SseK2348 SseK3335PhageST64B NleBwasidentifiedinC.rodentiumtogetherwithotherpotentialtypeIIIeffectorsencoded outsidetheLEE.TheyweredesignatedNleA,NleB,NleC,NleD,NleE,NleF,andNleGfornon‐LEE‐ encodedeffectors[106].TwohomologsofNleBareencodedinthegenomesofEHECandEPEC: NleB1andNleB2[106,108,109].NleB1is89%identicaltoNleBfromC.rodentium,andthereisa differenceinsevenaminoacidsbetweentheEHECandEPECversions.NleB2isidenticalinbothE. colipathotypesbutisonly60%identicaltoC.rodentiumNleB,althoughanorthologofnleB2ispresent inthegenomeofC.rodentiumasapseudogene[107].Interestingly,nleB2isanon‐neutralevolving gene,probablyduetopositiveselection,suggestingthatthisgeneisstillintheprocessofoptimizing itsfunction[114].S.entericastrainsencodethreehomologsofNleB:SseK1,SseK2,andSseK3 [111,112].Althoughthegenesencodingtheseeffectorsarenotlocatedinthepathogenicityislands wherethecognateT3SSareencoded,allofthemhaveevidenceofhorizontaltransferandsomeare associatedwithotherpathogenicityislandsorprophages(Table2).Interestingly,whereassseK1and sseK2arepresentinmostavailableSalmonellagenomesequences,thephage‐encodedSseK3effector hasalimiteddistribution[111].ABLASTsearchoncompletegenomesavailableattheNational CenterforBiotechnologyInformation(NCBI)revealsthatproteinsidenticalorverysimilartoNleB1 orNleB2areencodedindifferentstrainsoftheemergingA/EpathogenEscherichiaalbertii[115],and significantlysimilarproteinsarefoundinEscherichiamarmotae[116]andYersiniahibernica[117]. PercentagesofidentitybetweenrepresentativeNleB/SseKproteinsareshowninTable3,anda phylogeneticanalysisisshowninFigure4. Table3.PercentidentitydataforNleB/SseKproteinsprovidedbyClustalW. NleBC. rodentium NleB1 EPEC NleB1 EHEC NleBE. albertii NleBE. marmotaeNleB2SseK1SseK2SseK3 NleBC.rodentium NleB1EPEC88.75 NleB1EHEC89.0697.87 NleB1E.albertii88.4597.5799.09 NleBE.marmotae71.4271.7371.7371.43 NleB2EPEC/EHEC59.8261.0461.3560.4360.43 SseK1S.enterica57.4558.0559.5758.9758.9753.68 SseK2S.enterica52.5852.2853.1952.5851.0646.9355.36
Microorganisms2020,8,3579of23 SseK3S.enterica51.9853.1953.5052.8954.1047.5554.9375.22 NleBY.hibernica32.5232.8334.0434.0434.0433.1335.1233.1433.43 Figure4.PhylogeneticanalysisofNleB/SseKproteins.Alignmentandphylogeneticreconstructions wereperformedusingthefunction“build”ofETE3v3.1.1[118]asimplementedontheGenomeNet (https://www.genome.jp/tools/ete/).ThetreewasconstructedusingFastTreev2.1.8withdefault parameters[119].ValuesatnodesareSH‐likelocalsupport. 4.2.Expression,Translocation,andSubcellularLocalization EffectorsofT3SSsareexpectedtobeexpressedincoordinationwiththesynthesisoftheir cognatesecretionapparatus.ThisisthecaseforeffectorsoftheLEE,SPI1orSPI2systemsthatare encodedintheislands,buttheissueisnotsoobviousforeffectorsencodedoutsidetheislands, includingthemembersoftheNleB/SseKfamily. InA/Eorganisms,LeristhemaintranscriptionalregulatorofLEE.Itisencodedintheisland anditsleveliscontrolledbyothertranscriptionalregulatorsthatrespondtodifferentenvironmental stimuli,includinghosthormones,microbiota‐liberatedsugarsandmetabolites,molecularoxygenat thegutepithelium,andcontactofbacteriawithhostepithelialcells[120].Expressionisundetectable whenbacteriaaregrowninrichlaboratorymedia,butrapidexpressionandsecretionofproteinsto thesupernatantisobservedupontransfertoMEM‐HEPESmedium,whichisassociatedwiththe heterogeneousexpressionoftheEspADBtransloconapparatus[121].Severalnlegenes,including nleB,aretranscribedinEHECunderthesesecretion‐permissiveconditions,althoughinductionof nleBexpressionwasslight(twofold)andwasnotcoordinatedwiththesynthesisofEspAfilaments atthesingle‐celllevel[122].SecretionofNleB(NleB1inEPEC)bytheLEE‐encodedT3SShasbeen demonstratedinC.rodentiumandEPEC[106,110].TransfectionwithGFP‐fusionsortranslocationof TEM‐1β‐lactamasefusions[123]fromEPECintoHeLacellsrevealsadiffusecytosoliclocalizationof NleB1[110,124].TheregulatoryregionofEPECnleB2containsamotifconservedamongasubsetof genesencodingeffectorproteinsinA/EpathogensthatwasnamedNRIR(nleregulatoryinverted
Microorganisms2020,8,35716of23 Sequenceanalysisandevidenceforhorizontaltransferamongattachingandeffacingpathogens.Infect. Immun.2001,69,6323–6335. 21. Jarvis,K.G.;Girón,J.A.;Jerse,A.E.;Mcdaniel,T.K.;Donnenberg,M.S.;Kaper,J.B.Enteropathogenic EscherichiacolicontainsaputativetypeIIIsecretionsystemnecessaryfortheexportofproteinsinvolvedin attachingandeffacinglesionformation.Proc.Natl.Acad.Sci.USA1995,92,7996–8000. 22. Gaytán,M.O.;Martínez‐Santos,V.I.;Soto,E.;González‐Pedrajo,B.Typethreesecretionsysteminattaching andeffacingpathogens.Front.Cell.Infect.Microbiol.2016,6,129. 23. Zhou,M.;Guo,Z.;Duan,Q.;Hardwidge,P.R.;Zhu,G.EscherichiacolitypeIIIsecretionsystem2:Anew kindofT3SS?Vet.Res.2014,45,32. 24. Hayashi,T.;Makino,K.;Ohnishi,M.;Kurokawa,K.;Ishii,K.;Yokoyama,K.;Han,C.G.;Ohtsubo,E.; Nakayama,K.;Murata,T.;etal.CompletegenomesequenceofenterohemorrhagicEschelichiacoliO157:H7 andgenomiccomparisonwithalaboratorystrainK‐12.DNARes.2001,8,11–22. 25. Shulman,A.;Yair,Y.;Biran,D.;Sura,T.;Otto,A.;Gophna,U.;Becher,D.;Hecker,M.;Ron,E.Z.The EscherichiacolitypeIIIsecretionsystem2hasaglobaleffectoncellsurface.MBio2018,9,e01070‐18. 26. Ooka,T.;Ogura,Y.;Katsura,K.;Seto,K.;Kobayashi,H.;Kawano,K.;Tokuoka,E.;Furukawa,M.;Harada, S.;Yoshino,S.;etal.DefiningthegenomefeaturesofEscherichiaalbertii,anemergingenteropathogen closelyrelatedtoEscherichiacoli.GenomeBiol.Evol.2015,7,3170–3179. 27. Ideses,D.;Gophna,U.;Paitan,Y.;Chaudhuri,R.R.;Pallen,M.J.;Ron,E.Z.AdegeneratetypeIIIsecretion systemfromsepticemicEscherichiacolicontributestopathogenesis.J.Bacteriol.2005,187,8164–8171. 28. Hurley,D.;McCusker,M.P.;Fanning,S.;Martins,M.Salmonella‐hostinteractions—Modulationofthehost innateimmunesystem.Front.Immunol.2014,5,481. 29. Lairson,L.L.;Henrissat,B.;Davies,G.J.;Withers,S.G.Glycosyltransferases:Structures,functions,and mechanisms.Annu.Rev.Biochem.2008,77,521–555. 30. Mestrom,L.;Przypis,M.;Kowalczykiewicz,D.;Pollender,A.;Kumpf,A.;Marsden,S.R.;Bento,I.;Jarzębski, A.B.;Szymańska,K.;Chruściel,A.;etal.Leloirglycosyltransferasesinappliedbiocatalysis:A multidisciplinaryapproach.Int.J.Mol.Sci.2019,20,5263. 31. Campbell,J.A.;Davies,G.J.;Bulone,V.;Henrissat,B.Aclassificationofnucleotide‐diphospho‐sugar glycosyltransferasesbasedonaminoacidsequencesimilarities.Biochem.J.1997,326,929–939. 32. Coutinho,P.M.;Deleury,E.;Davies,G.J.;Henrissat,B.Anevolvinghierarchicalfamilyclassificationfor glycosyltransferases.J.Mol.Biol.2003,328,307–317. 33. Sinnott,M.L.Catalyticmechanismofenzymicglycosyltransfer.Chem.Rev.1990,90,1171–1202. 34. Liang,D.M.;Liu,J.H.;Wu,H.;Wang,B.Bin;Zhu,H.J.;Qiao,J.J.Glycosyltransferases:Mechanismsand applicationsinnaturalproductdevelopment.Chem.Soc.Rev.2015,44,8350–8374. 35. Davies,G.J.Sweetsecretsofsynthesis.Nat.Struct.Biol.2001,8,98–100. 36. Vocadlo,D.J.;Davies,G.J.;Laine,R.;Withers,S.G.Catalysisbyhenegg‐whitelysozymeproceedsviaa covalentintermediate.Nature2001,412,835–838. 37. Soya,N.;Fang,Y.;Palcic,M.M.;Klassen,J.S.Trappingandcharacterizationofcovalentintermediatesof mutantretainingglycosyltransferases.Glycobiology2011,21,547–552. 38. Lira‐Navarrete,E.;Iglesias‐Fernández,J.;Zandberg,W.F.;Compañón,I.;Kong,Y.;Corzana,F.;Pinto,B.M.; Clausen,H.;Peregrina,J.M.;Vocadlo,D.J.;etal.Substrate‐guidedfront‐facereactionrevealedbycombined structuralsnapshotsandmetadynamicsforthepolypeptideN‐acetylgalactosaminyltransferase2.Angew. Chem.Int.Ed.Engl.2014,53,8206–8210. 39. Ardèvol,A.;Rovira,C.Themolecularmechanismofenzymaticglycosyltransferwithretentionof configuration:Evidenceforashort‐livedoxocarbenium‐likespecies.Angew.Chem.Int.Ed.2011,50,10897– 10901. 40. Schuman,B.;Evans,S.V;Fyles,T.M.Geometricattributesofretainingglycosyltransferaseenzymesfavor anorthogonalmechanism.PLoSONE2013,8,e71077. 41. Dodson,E.;Harding,M.M.;Hodgkin,D.C.;Rossmann,M.G.Thecrystalstructureofinsulin:III.Evidence fora2‐foldaxisinrhombohedralzincinsulin.J.Mol.Biol.1966,16,227–241. 42. Schuman,B.;Alfaro,J.A.;Evans,S.V.GlycosyltransferaseStructureandFunction.InBioactiveConformation I;Springer:Berlin/Heidelberg,Germany,2006;pp.217–257. 43. Kikuchi,N.;Kwon,Y.D.;Gotoh,M.;Narimatsu,H.Comparisonofglycosyltransferasefamiliesusingthe profilehiddenMarkovmodel.Biochem.Biophys.Res.Commun.2003,310,574–579. 44. Liu,J.;Mushegian,A.Threemonophyleticsuperfamiliesaccountforthemajorityoftheknown
Microorganisms2020,8,35717of23 glycosyltransferases.ProteinSci.2003,12,1418–1431. 45. Rosén,M.L.;Edman,M.;Sjöström,M.;Wieslander,A.Recognitionoffoldandsugarlinkagefor glycosyltransferasesbymultivariatesequenceanalysis.J.Biol.Chem.2004,279,38683–38692. 46. Lizak,C.;Gerber,S.;Numao,S.;Aebi,M.;Locher,K.P.X‐raystructureofabacterial oligosaccharyltransferase.Nature2011,474,350–356. 47. Zhang,H.;Zhu,F.;Yang,T.;Ding,L.;Zhou,M.;Li,J.;Haslam,S.M.;Dell,A.;Erlandsen,H.;Wu,H.The highlyconserveddomainofunknownfunction1792hasadistinctglycosyltransferasefold.Nat.Commun. 2014,5,4339. 48. Lovering,A.L.;DeCastro,L.H.;Lim,D.;Strynadka,N.C.J.Structuralinsightintothetransglycosylation stepofbacterialcell‐wallbiosynthesis.Science(80‐)2007,315,1402–1405. 49. Yuan,Y.;Barrett,D.;Zhang,Y.;Kahne,D.;Sliz,P.;Walker,S.Crystalstructureofapeptidoglycan glycosyltransferasesuggestsamodelforprocessiveglycanchainsynthesis.Proc.Natl.Acad.Sci.USA2007, 104,5348–5353. 50. Kattke,M.D.;Gosschalk,J.E.;Martinez,O.E.;Kumar,G.;Gale,R.T.;Cascio,D.;Sawaya,M.R.;Philips,M.; Brown,E.D.;Clubb,R.T.StructureandmechanismofTagA,anovelmembrane‐associated glycosyltransferasethatproduceswallteichoicacidsinpathogenicbacteria.PLoSPathog.2019,15,e1007723. 51. Charnock,S.J.;Davies,G.J.Structureofthenucleotide‐diphospho‐sugartransferase,SpsAfromBacillus subtilis,innativeandnucleotide‐complexedforms.Biochemistry1999,38,6380–6385. 52. Mulichak,A.M.;Losey,H.C.;Walsh,C.T.;Garavito,R.M.StructureoftheUDP‐glucosyltransferaseGtfB thatmodifiestheheptapeptideaglyconeinthebiosynthesisofvancomycingroupantibiotics.Structure2001, 9,547–557. 53. Rose,A.S.;Bradley,A.R.;Valasatava,Y.;Duarte,J.M.;Prlić,A.;Rose,P.W.NGLviewer:Web‐based moleculargraphicsforlargecomplexes.Bioinformatics2018,34,3755–3758. 54. Lu,Q.;Li,S.;Shao,F.Sweettalk:Proteinglycosylationinbacterialinteractionwiththehost.TrendsMicrobiol. 2015,23,630–641. 55. Nothaft,H.;Szymanski,C.M.Proteinglycosylationinbacteria:Sweeterthanever.Nat.Rev.Microbiol.2010, 8,765–778. 56. Grass,S.;Buscher,A.Z.;Swords,W.E.;Apicella,M.A.;Barenkamp,S.J.;Ozchlewski,N.;StGeme,J.W.The HaemophilusinfluenzaeHMW1adhesinisglycosylatedinaprocessthatrequiresHMW1Cand phosphoglucomutase,anenzymeinvolvedinlipooligosaccharidebiosynthesis.Mol.Microbiol.2003,48, 737–751. 57. StGeme,J.W.;Grass,S.SecretionoftheHaemophilusinfluenzaeHMW1andHMW2adhesinsinvolvesa periplasmicintermediateandrequirestheHMWBandHMWCproteins.Mol.Microbiol.1998,27,617–630. 58. Gross,J.;Grass,S.;Davis,A.E.;Gilmore‐Erdmann,P.;Townsend,R.R.;St.Geme,J.W.TheHaemophilus influenzaeHMW1adhesinisaglycoproteinwithanunusualN‐linkedcarbohydratemodification.J.Biol. Chem.2008,283,26010–26015. 59. Szymanski,C.M.;Burr,D.H.;Guerry,P.Campylobacterproteinglycosylationaffectshostcellinteractions. Infect.Immun.2002,70,2242–2244. 60. Karlyshev,A.V.;Everest,P.;Linton,D.;Cawthraw,S.;Newell,D.G.;Wren,B.W.TheCampylobacterjejuni generalglycosylationsystemisimportantforattachmenttohumanepithelialcellsandinthecolonization ofchicks.Microbiology2004,150,1957–1964. 61. Hendrixson,D.R.;DiRita,V.J.IdentificationofCampylobacterjejunigenesinvolvedincommensal colonizationofthechickgastrointestinaltract.Mol.Microbiol.2004,52,471–484. 62. Perez,C.;Gerber,S.;Boilevin,J.;Bucher,M.;Darbre,T.;Aebi,M.;Reymond,J.‐L.;Locher,K.P.Structure andmechanismofanactivelipid‐linkedoligosaccharideflippase.Nature2015,524,433–438. 63. Wacker,M.;Linton,D.;Hitchen,P.G.;Nita‐Lazar,M.;Haslam,S.M.;North,S.J.;Panico,M.;Morris,H.R.; Dell,A.;Wren,B.W.;etal.N‐linkedglycosylationinCampylobacterjejunianditsfunctionaltransferintoE. coli.Science2002,298,1790–1793. 64. Bokhari,H.;Maryam,A.;Shahid,R.;Siddiqi,A.R.OligosaccharyltransferasePglBofCampylobacterjejuniis aglycoprotein.WorldJ.Microbiol.Biotechnol.2020,36,9. 65. Lu,Q.;Yao,Q.;Xu,Y.;Li,L.;Li,S.;Liu,Y.;Gao,W.;Niu,M.;Sharon,M.;Ben‐Nissan,G.;etal.Anironcontainingdodecamericheptosyltransferasefamilymodifiesbacterialautotransportersinpathogenesis. CellHostMicrobe2014,16,351–363. 66. Benz,I.;Schmidt,M.A.Glycosylationwithheptoseresiduesmediatedbytheaahgeneproductisessential
Microorganisms2020,8,35718of23 foradherenceoftheAIDA‐Iadhesin.Mol.Microbiol.2001,40,1403–1413. 67. Zhou,M.;Wu,H.Glycosylationandbiogenesisofafamilyofserine‐richbacterialadhesins.Microbiology 2009,155,317–327. 68. Lizcano,A.;Sanchez,C.J.;Orihuela,C.J.Aroleforglycosylatedserine‐richrepeatproteinsingram‐positive bacterialpathogenesis.Mol.OralMicrobiol.2012,27,257–269. 69. Zhu,F.;Zhang,H.;Yang,T.;Haslam,S.M.;Dell,A.;Wu,H.Engineeringanddissectingtheglycosylation pathwayofastreptococcalserine‐richrepeatadhesin.J.Biol.Chem.2016,291,27354–27363. 70. Logan,S.M.Flagellarglycosylation—Anewcomponentofthemotilityrepertoire?Microbiology2006,152, 1249–1262. 71. Duan,Q.;Zhou,M.;Zhu,L.;Zhu,G.Flagellaandbacterialpathogenicity.J.BasicMicrobiol.2013,53,1–8. 72. Merino,S.;Tomás,J.M.Gram‐negativeflagellaglycosylation.Int.J.Mol.Sci.2014,15,2840–2857. 73. Delvaux,N.A.;Thoden,J.B.;Holden,H.M.MoleculararchitecturesofPenandPal:Keyenzymesrequired forCMP‐pseudaminicacidbiosynthesisinBacillusthuringiensis.ProteinSci.2018,27,738–749. 74. Valiente,E.;Bouché,L.;Hitchen,P.;Faulds‐Pain,A.;Songane,M.;Dawson,L.F.;Donahue,E.;Stabler,R.A.; Panico,M.;Morris,H.R.;etal.RoleofglycosyltransferasesmodifyingtypeBflagellinofemerging hypervirulentClostridiumdifficilelineagesandtheirimpactonmotilityandbiofilmformation.J.Biol.Chem. 2016,291,25450–25461. 75. Bouché,L.;Panico,M.;Hitchen,P.;Binet,D.;Sastre,F.;Faulds‐Pain,A.;Valiente,E.;Vinogradov,E.;Aubry, A.;Fulton,K.;etal.ThetypeBflagellinofhypervirulentClostridiumdifficileismodifiedwithnovel sulfonatedpeptidylamido‐glycans.J.Biol.Chem.2016,291,25439–25449. 76. DeMaayer,P.;Cowan,D.A.Comparativegenomicanalysisoftheflagellinglycosylationislandofthe Gram‐positivethermophileGeobacillus.BMCGenom.2016,17,913. 77. Janesch,B.;Schirmeister,F.;Maresch,D.;Altmann,F.;Messner,P.;Kolarich,D.;Schäffer,C.Flagellin glycosylationinPaenibacillusalveiCCM2051T.Glycobiology2016,26,74–87. 78. Twine,S.M.;Reid,C.W.;Aubry,A.;McMullin,D.R.;Fulton,K.M.;Austin,J.;Logan,S.M.Motilityand flagellarglycosylationinClostridiumdifficile.J.Bacteriol.2009,191,7050–7062. 79. Twine,S.M.;Paul,C.J.;Vinogradov,E.;McNally,D.J.;Brisson,J.R.;Mullen,J.A.;McMullin,D.R.;Jarrell, H.C.;Austin,J.W.;Kelly,J.F.;etal.FlagellarglycosylationinClostridiumbotulinum.FEBSJ.2008,275,4428– 4444. 80. Schirm,M.;Kalmokoff,M.;Aubry,A.;Thibault,P.;Sandoz,M.;Logan,S.M.FlagellinfromListeria monocytogenesisglycosylatedwithβ‐O‐linkedN‐acetylglucosamine.J.Bacteriol.2004,186,6721–6727. 81. Haya,S.;Tokumaru,Y.;Abe,N.;Kaneko,J.;Aizawa,S.I.CharacterizationoflateralflagellaofSelenomonas ruminantium.Appl.Environ.Microbiol.2011,77,2799–2802. 82. Merino,S.;Aquilini,E.;Fulton,K.M.;Twine,S.M.;Tomás,J.M.Thepolarandlateralflagellafrom Plesiomonasshigelloidesareglycosylatedwithlegionaminicacid.Front.Microbiol.2015,6,649. 83. Scott,A.E.;Twine,S.M.;Fulton,K.M.;Titball,R.W.;Essex‐Lopresti,A.E.;Atkins,T.P.;Prior,J.L.Flagellar glycosylationinBurkholderiapseudomalleiandBurkholderiathailandensis.J.Bacteriol.2011,193,3577–3587. 84. Rath,C.B.;Schirmeister,F.;Figl,R.;Seeberger,P.H.;Schäffer,C.;Kolarich,D.Flagellinglycoproteomicsof theperiodontitisassociatedpathogenSelenomonassputigenarevealspreviouslynotdescribedO‐glycans andrhamnosefragmentrearrangementoccurringontheglycopeptides.Mol.Cell.Proteom.2018,17,721– 736. 85. Thibault,P.;Logan,S.M.;Kelly,J.F.;Brisson,J.R.;Ewing,C.P.;Trust,T.J.;Guerry,P.Identificationofthe carbohydratemoietiesandglycosylationmotifsinCampylobacterjejuniflagellin.J.Biol.Chem.2001,276, 34862–34870. 86. Parker,J.L.;Day‐Williams,M.J.;Tomas,J.M.;Stafford,G.P.;Shaw,J.G.Identificationofaputative glycosyltransferaseresponsibleforthetransferofpseudaminicacidontothepolarflagellinofAeromonas caviaeSch3N.Microbiologyopen2012,1,149–160. 87. Rabaan,A.A.;Gryllos,I.;Tomás,J.M.;Shaw,J.G.Motilityandthepolarflagellumarerequiredfor AeromonascaviaeadherencetoHEp‐2cells.Infect.Immun.2001,69,4257–4267. 88. Wilhelms,M.;Fulton,K.M.;Twine,S.M.;Tomás,J.M.;Merino,S.Differentialglycosylationofpolarand lateralflagellinsinAeromonashydrophilaAH‐3.J.Biol.Chem.2012,287,27851–27862. 89. Dorrell,N.;Mangan,J.A.;Laing,K.G.;Hinds,J.;Linton,D.;Al‐Ghusein,H.;Barrell,B.G.;Parkhill,J.;Stoker, N.G.;Karlyshev,A.V.;etal.WholegenomecomparisonofCampylobacterjejunihumanisolatesusingalowcostmicroarrayrevealsextensivegeneticdiversity.GenomeRes.2001,11,1706–1715.
Microorganisms2020,8,35719of23 90. Alm,R.A.;Guerry,P.;Power,M.E.;Trust,T.J.Variationinantigenicityandmolecularweightof CampylobactercoliVC167flagellinindifferentgeneticbackgrounds.J.Bacteriol.1992,174,4230–4238. 91. Schoenhofen,I.C.;McNally,D.J.;Brisson,J.‐R.;Logan,S.M.ElucidationoftheCMP‐pseudaminicacid pathwayinHelicobacterpylori:SynthesisfromUDP‐N‐acetylglucosaminebyasingleenzymaticreaction. Glycobiology2006,16,8C–14C. 92. Schirm,M.;Soo,E.C.;Aubry,A.J.;Austin,J.;Thibault,P.;Logan,S.M.Structural,geneticandfunctional characterizationoftheflagellinglycosylationprocessinHelicobacterpylori.Mol.Microbiol.2003,48,1579– 1592. 93. Sulzenbacher,G.;Roig‐Zamboni,V.;Lebrun,R.;Guérardel,Y.;Murat,D.;Mansuelle,P.;Yamakawa,N.; Qian,X.X.;Vincentelli,R.;Bourne,Y.;etal.Glycosylateandmove!TheglycosyltransferaseMafisinvolved inbacterialflagellaformation.Environ.Microbiol.2018,20,228–240. 94. McNally,D.J.;Hui,J.P.M.;Aubry,A.J.;Mui,K.K.K.;Guerry,P.;Brisson,J.R.;Logan,S.M.;Soo,E.C. FunctionalcharacterizationoftheflagellarglycosylationlocusinCampylobacterjejuni81–176usinga focusedmetabolomicsapproach.J.Biol.Chem.2006,281,18489–18498. 95. Parker,J.L.;Lowry,R.C.;Couto,N.A.S.;Wright,P.C.;Stafford,G.P.;Shaw,J.G.Maf‐dependentbacterial flagellinglycosylationoccursbeforechaperonebindingandflagellarT3SSexport.Mol.Microbiol.2014,92, 258–272. 96. Voth,D.E.;Ballard,J.D.Clostridiumdifficiletoxins:Mechanismofactionandroleindisease.Clin.Microbiol. Rev.2005,18,247–263. 97. Kuehne,S.A.;Cartman,S.T.;Heap,J.T.;Kelly,M.L.;Cockayne,A.;Minton,N.P.TheroleoftoxinAand toxinBinClostridiumdifficileinfection.Nature2010,467,711–713. 98. Just,I.;Selzer,J.;Wilm,M.;vonEichel‐Streiber,C.;Mann,M.;Aktories,K.GlucosylationofRhoproteins byClostridiumdifficiletoxinB.Nature1995,375,500–503. 99. Genth,H.;Aktories,K.;Just,I.MonoglucosylationofRhoAatthreonine37blockscytosol‐membrane cycling.J.Biol.Chem.1999,274,29050–29056. 100. Jank,T.;Bogdanović,X.;Wirth,C.;Haaf,E.;Spoerner,M.;Böhmer,K.E.;Steinemann,M.;Orth,J.H.C.; Kalbitzer,H.R.;Warscheid,B.;etal.AbacterialtoxincatalyzingtyrosineglycosylationofRhoand deamidationofGqandGiproteins.Nat.Struct.Mol.Biol.2013,20,1273–1280. 101. Qiu,J.;Luo,Z.‐Q.LegionellaandCoxiellaeffectors:Strengthindiversityandactivity.Nat.Rev.Microbiol. 2017,15,591–605. 102. Belyi,Y.;Tabakova,I.;Stahl,M.;Aktories,K.Lgt:Afamilyofcytotoxicglucosyltransferasesproducedby Legionellapneumophila.J.Bacteriol.2008,190,3026–3035. 103. Lü,W.;Du,J.;Stahl,M.;Tzivelekidis,T.;Belyi,Y.;Gerhardt,S.;Aktories,K.;Einsle,O.Structuralbasisof theactionofglucosyltransferaseLgt1fromLegionellapneumophila.J.Mol.Biol.2010,396,321–331. 104. Reinert,D.J.;Jank,T.;Aktories,K.;Schulz,G.E.StructuralbasisforthefunctionofClostridiumdifficiletoxin B.J.Mol.Biol.2005,351,973–981. 105. Belyi,Y.;Stahl,M.;Sovkova,I.;Kaden,P.;Luy,B.;Aktories,K.Regionofelongationfactor1A1involved insubstraterecognitionbyLegionellapneumophilaglucosyltransferaseLgt1.IdentificationofLgt1asa retainingglucosyltransferase.J.Biol.Chem.2009,284,20167–20174. 106. Deng,W.;Puente,J.L.;Gruenheid,S.;Li,Y.;Vallance,B.A.;Vázquez,A.;Barba,J.;Ibarra,J.A.;O’Donnell, P.;Metalnikov,P.;etal.Dissectingvirulence:Systematicandfunctionalanalysesofapathogenicityisland. Proc.Natl.Acad.Sci.USA2004,101,3597–3602. 107. Petty,N.K.;Bulgin,R.;Crepin,V.F.;Cerdeño‐Tárraga,A.M.;Schroeder,G.N.;Quail,M.A.;Lennard,N.; Corton,C.;Barron,A.;Clark,L.;etal.TheCitrobacterrodentiumgenomesequencerevealsconvergent evolutionwithhumanpathogenicEscherichiacoli.J.Bacteriol.2010,192,525–538. 108. Perna,N.T.;Plunkett,G.;Burland,V.;Mau,B.;Glasner,J.D.;Rose,D.J.;Mayhew,G.F.;Evans,P.S.;Gregor, J.;Kirkpatrick,H.A.;etal.GenomesequenceofenterohaemorrhagicEscherichiacoliO157:H7.Nature2001, 409,529–533. 109. Iguchi,A.;Thomson,N.R.;Ogura,Y.;Saunders,D.;Ooka,T.;Henderson,I.R.;Harris,D.;Asadulghani,M.; Kurokawa,K.;Dean,P.;etal.Completegenomesequenceandcomparativegenomeanalysisof enteropathogenicEscherichiacoliO127:H6strainE2348/69.J.Bacteriol.2009,91,347–354. 110. Kelly,M.;Hart,E.;Mundy,R.;Marchès,O.;Wiles,S.;Badea,L.;Luck,S.;Tauschek,M.;Frankel,G.;Robins‐ Browne,R.M.;etal.EssentialroleofthetypeIIIsecretionsystemeffectorNleBincolonizationofmiceby Citrobacterrodentium.Infect.Immun.2006,74,2328–2337.
Microorganisms2020,8,35720of23 111. Brown,N.F.;Coombes,B.K.;Bishop,J.L.;Wickham,M.E.;Lowden,M.J.;Gal‐Mor,O.;Goode,D.L.;Boyle, E.C.;Sanderson,K.L.;Finlay,B.B.;etal.SalmonellaphageST64BencodesamemberoftheSseK/NleB effectorfamily.PLoSONE2011,6,e17824. 112. KujatChoy,S.L.;Boyle,E.C.;Gal‐Mor,O.;Goode,D.L.;Valdez,Y.;Vallance,B.A.;Finlay,B.B.SseK1and SseK2arenoveltranslocatedproteinsofSalmonellaentericaserovarTyphimurium.Infect.Immun.2004,72, 5115–5125. 113. Jarvik,T.;Smillie,C.;Groisman,E.A.;Ochman,H.Short‐termsignaturesofevolutionarychangeinthe SalmonellaentericaserovarTyphimurium14028genome.J.Bacteriol.2010,192,560–567. 114. Eswarappa,S.M.;Janice,J.;Balasundaram,S.V;Chakravortty,D.Non‐neutralevolutioninnon‐LEE‐ encodedtypeIIIeffectorsofattachingandeffacingEscherichiacoli.MicrobesInfect.2013,15,147–151. 115. Bhatt,S.;Egan,M.;Critelli,B.;Kouse,A.;Kalman,D.;Upreti,C.Theevasiveenemy:Insightsintothe virulenceandepidemiologyoftheemergingattachingandeffacingpathogenEscherichiaalbertii.Infect. Immun.2019,87,e00254‐18. 116. Liu,S.;Feng,J.;Pu,J.;Xu,X.;Lu,S.;Yang,J.;Wang,Y.;Jin,D.;Du,X.;Meng,X.;etal.Genomicand molecularcharacterisationofEscherichiamarmotaefromwildrodentsinQinghai‐Tibetplateauasapotential pathogen.Sci.Rep.2019,9,10619. 117. Nguyen,S.V.;Muthappa,D.M.;Hurley,D.;Donoghue,O.;McCabe,E.;Anes,J.;Schaffer,K.;Murphy,B.P.; Buckley,J.F.;Fanning,S.Yersiniahibernicasp.nov.,isolatedfrompig‐productionenvironments.Int.J.Syst. Evol.Microbiol.2019,69,2023–2027. 118. Huerta‐Cepas,J.;Serra,F.;Bork,P.ETE3:Reconstruction,analysis,andvisualizationofphylogenomicdata. Mol.Biol.Evol.2016,33,1635–1638. 119. Price,M.N.;Dehal,P.S.;Arkin,A.P.Fasttree:Computinglargeminimumevolutiontreeswithprofiles insteadofadistancematrix.Mol.Biol.Evol.2009,26,1641–1650. 120. Hughes,D.T.;Sperandio,V.Inter‐kingdomsignalling:Communicationbetweenbacteriaandtheirhosts. Nat.Rev.Microbiol.2008,6,111–120. 121. Roe,A.J.;Yull,H.;Naylor,S.W.;Woodward,M.J.;Smith,D.G.E.;Gally,D.L.Heterogeneoussurface expressionofEspAtransloconfilamentsbyEscherichiacoliO157:H7iscontrolledattheposttranscriptional level.Infect.Immun.2003,71,5900–5909. 122. Roe,A.J.;Tysall,L.;Dransfield,T.;Wang,D.;Fraser‐Pitt,D.;Mahajan,A.;Constandinou,C.;Inglis,N.; Downing,A.;Talbot,R.;etal.Analysisoftheexpression,regulationandexportofNleA‐EinEscherichiacoli O157:H7.Microbiology2007,153,1350–1360. 123. Charpentier,X.;Oswald,E.Identificationofthesecretionandtranslocationdomainoftheenteropathogenic andenterohemorrhagicEscherichiacolieffectorCif,usingTEM‐1beta‐lactamaseasanewfluorescencebasedreporter.J.Bacteriol.2004,186,5486–5495. 124. Newton,H.J.;Pearson,J.S.;Badea,L.;Kelly,M.;Lucas,M.;Holloway,G.;Wagstaff,K.M.;Dunstone,M.A.; Sloan,J.;Whisstock,J.C.;etal.ThetypeIIIeffectorsNleEandNleBfromenteropathogenicE.coliandOspZ fromShigellablocknucleartranslocationofNF‐kappaBp65.PLoSPathog.2010,6,e1000898. 125. García‐Angulo,V.A.;Martínez‐Santos,V.I.;Villaseñor,T.;Santana,F.J.;Huerta‐Saquero,A.;Martínez,L.C.; Jiménez,R.;Lara‐Ochoa,C.;Téllez‐Sosa,J.;Bustamante,V.H.;etal.Adistinctregulatorysequenceis essentialfortheexpressionofasubsetofnlegenesinattachingandeffacingEscherichiacoli.J.Bacteriol.2012, 194,5589–5603. 126. Deng,W.;Yu,H.B.;DeHoog,C.L.;Stoynov,N.;Li,Y.;Foster,L.J.;Finlay,B.B.Quantitativeproteomic analysisoftypeIIIsecretomeofenteropathogenicEscherichiacolirevealsanexpandedeffectorrepertoire forattaching/effacingbacterialpathogens.Mol.Cell.Proteom.2012,11,692–709. 127. Ellermeier,C.D.;Ellermeier,J.R.;Slauch,J.M.HilD,HilCandRtsAconstituteafeedforwardloopthat controlsexpressionoftheSPI1typethreesecretionsystemregulatorhilAinSalmonellaentericaserovar Typhimurium.Mol.Microbiol.2005,57,691–705. 128. Schechter,L.M.;Jain,S.;Akbar,S.;Lee,C.A.Thesmallnucleoid‐bindingproteinsH‐NS,HU,andFisaffect hilAexpressioninSalmonellaentericaserovarTyphimurium.Infect.Immun.2003,71,5432–5435. 129. Laughlin,R.C.;Knodler,L.A.;Barhoumi,R.;RossPayne,H.;Wu,J.;Gomez,G.;Pugh,R.;Lawhon,S.D.; Bäumler,A.J.;Steele‐Mortimer,O.;etal.Spatialsegregationofvirulencegeneexpressionduringacute entericinfectionwithSalmonellaentericaserovarTyphimurium.MBio2014,5,e00946‐13. 130. Mouslim,C.;Delgado,M.;Groisman,E.A.ActivationoftheRcsC/YojN/RcsBphosphorelaysystem attenuatesSalmonellavirulence.Mol.Microbiol.2004,54,386–395.
Microorganisms2020,8,35721of23 131. Golubeva,Y.A.;Sadik,A.Y.;Ellermeier,J.R.;Slauch,J.M.Integratingglobalregulatoryinputintothe SalmonellapathogenicityIsland1typeIIIsecretionsystem.Genetics2012,190,79–90. 132. Palmer,A.D.;Kim,K.;Slauch,J.M.PhoP‐mediatedrepressionoftheSPI1T3SSinSalmonellaentericaserovar Typhimurium.J.Bacteriol.2019,201,e00264‐19. 133. Prost,L.R.;Miller,S.I.TheSalmonellaePhoQsensor:Mechanismsofdetectionofphagosomesignals.Cell. Microbiol.2008,10,576–582. 134. Kato,A.;Groisman,E.A.;HowardHughesMedicalInstituteThePhoQ/PhoPregulatorynetworkof Salmonellaenterica.Adv.Exp.Med.Biol.2008,631,7–21. 135. Bijlsma,J.J.E.;Groisman,E.A.ThePhoP/PhoQsystemcontrolstheintramacrophagetypethreesecretion systemofSalmonellaenterica.Mol.Microbiol.2005,57,85–96. 136. Baisón‐Olmo,F.;Galindo‐Moreno,M.;Ramos‐Morales,F.Hostcelltype‐dependenttranslocationand PhoP‐mediatedpositiveregulationoftheeffectorSseK1ofSalmonellaenterica.Front.Microbiol.2015,6,396. 137. Günster,R.A.;Matthews,S.A.;Holden,D.W.;Thurston,T.L.M.SseK1andSseK3T3SSeffectorsinhibitNF‐ κBsignallingandnecroptoticcelldeathinSalmonella‐infectedmacrophages.Infect.Immun.2017, doi:10.1128/IAI.00010‐17. 138. Yang,Z.;Soderholm,A.;Lung,T.W.F.;Giogha,C.;Hill,M.M.;Brown,N.F.;Hartland,E.;Teasdale,R.D. SseK3IsaSalmonellaeffectorthatbindsTRIM32andmodulatesthehost’sNF‐κBsignallingactivity.PLoS ONE2015,10,e0138529. 139. Singh,D.G.;Lomako,J.;Lomako,W.M.;Whelan,W.J.;Meyer,H.E.;Serwe,M.;Metzger,J.W.beta‐ Glucosylarginine:anewglucose‐proteinbondinaself‐glucosylatingproteinfromsweetcorn.FEBSLett. 1995,376,61–64. 140. Li,S.;Zhang,L.;Yao,Q.;Li,L.;Dong,N.;Rong,J.;Gao,W.;Ding,X.;Sun,L.;Chen,X.;etal.Pathogen blockshostdeathreceptorsignallingbyarginineGlcNAcylationofdeathdomains.Nature2013,501,242– 246. 141. Pearson,J.S.;Giogha,C.;Ong,S.Y.;Kennedy,C.L.;Kelly,M.;Robinson,K.S.;Lung,T.W.F.;Mansell,A.; Riedmaier,P.;Oates,C.V.L.;etal.AtypeIIIeffectorantagonizesdeathreceptorsignallingduringbacterial gutinfection.Nature2013,501,247–251. 142. Gao,X.;Pham,T.H.;Feuerbacher,L.A.;Chen,K.;Hays,M.P.;Singh,G.;Rueter,C.;Hurtado‐Guerrero,R.; Hardwidge,P.R.CitrobacterrodentiumNleBproteininhibitsTumorNecrosisFactor(TNF)receptorassociatedfactor3(TRAF3)ubiquitinationtoreducehosttypeIinterferonproduction.J.Biol.Chem.2016, 291,18232–18238. 143. ElQaidi,S.;Chen,K.;Halim,A.;Siukstaite,L.;Rueter,C.;Hurtado‐Guerrero,R.;Clausen,H.;Hardwidge, P.R.NleB/SseKeffectorsfromCitrobacterrodentium,Escherichiacoli,andSalmonellaentericadisplaydistinct differencesinhostsubstratespecificity.J.Biol.Chem.2017,292,11423–11430. 144. Esposito,D.;Günster,R.A.;Martino,L.;ElOmari,K.;Wagner,A.;Thurston,T.L.M.;Rittinger,K.Structural basisfortheglycosyltransferaseactivityoftheSalmonellaeffectorSseK3.J.Biol.Chem.2018,293,5064–5078. 145. Park,J.B.;Kim,Y.H.;Yoo,Y.;Kim,J.;Jun,S.‐H.;Cho,J.W.;ElQaidi,S.;Walpole,S.;Monaco,S.;García‐ García,A.A.;etal.Structuralbasisforarginineglycosylationofhostsubstratesbybacterialeffectorproteins. Nat.Commun.2018,9,4283. 146. Ding,J.;Pan,X.;Du,L.;Yao,Q.;Xue,J.;Yao,H.;Wang,D.‐C.;Li,S.;Shao,F.Structuralandfunctional insightsintohostdeathdomainsinactivationbythebacterialarginineGlcNAcyltransferaseeffector.Mol. Cell2019,74,922–935.e6. 147. Newson,J.P.;Scott,N.E.;YeukWahChung,I.;WongFokLung,T.;Giogha,C.;Gan,J.;Wang,N.;Strugnell, R.;Brown,N.F.;Cygler,M.;etal.SalmonellaeffectorsSseK1andSseK3targetdeathdomainproteinsinthe TNFandTRAILsignalingpathways.Mol.Cell.Proteom.2019,doi:10.1074/mcp.RA118.001093. 148. Pan,M.;Li,S.;Li,X.;Shao,F.;Liu,L.;Hu,H.‐G.Synthesisofandspecificantibodygenerationfor glycopeptideswitharginineN‐GlcNAcylation.Angew.Chem.Int.Ed.Engl.2014,53,14517–14521. 149. Park,J.B.;Yoo,Y.;Cho,H.‐S.Structuralinsightsshowinghowarginineisabletobeglycosylatedby pathogeniceffectorproteins.BMBRep.2018,51,609–610. 150. Ghosh,S.;O’Connor,T.J.Beyondparalogs:Themultiplelayersofredundancyinbacterialpathogenesis. Front.Cell.Infect.Microbiol.2017,7,467. 151. Gao,X.;Wang,X.;Pham,T.H.;Feuerbacher,L.A.;Lubos,M.‐L.;Huang,M.;Olsen,R.;Mushegian,A.; Slawson,C.;Hardwidge,P.R.NleB,abacterialeffectorwithglycosyltransferaseactivity,targetsGAPDH functiontoinhibitNF‐κBactivation.CellHostMicrobe2013,13,87–99.
Microorganisms2020,8,35722of23 152. WongFokLung,T.;Giogha,C.;Creuzburg,K.;Ong,S.Y.;Pollock,G.L.;Zhang,Y.;Fung,K.Y.;Pearson,J.S.; Hartland,E.L.Mutagenesisandfunctionalanalysisofthebacterialarginineglycosyltransferaseeffector NleB1fromenteropathogenicEscherichiacoli.Infect.Immun.2016,84,1346–1360. 153. Cornick,N.A.;Pitzer,J.;Helgerson,A.F.;Madsen,M.L.;Kurth,K.T.;Xiao,Q.;Minion,F.C.UseofsignaturetaggedmutagenesistoidentifygenesassociatedwithcolonizationofsheepbyE.coliO157:H7.Vet.Microbiol. 2017,201,177–182. 154. Misyurina,O.;Asper,D.J.;Deng,W.;Finlay,B.B.;Rogan,D.;Potter,A.A.TheroleofTir,EspA,andNleB inthecolonizationofcattlebyShigatoxinproducingEscherichiacoliO26:H11.Can.J.Microbiol.2010,56, 739–747. 155. Afset,J.E.;Bruant,G.;Brousseau,R.;Harel,J.;Anderssen,E.;Bevanger,L.;Bergh,K.Identificationof virulencegeneslinkedwithdiarrheaduetoatypicalenteropathogenicEscherichiacolibyDNAmicroarray analysisandPCR.J.Clin.Microbiol.2006,44,3703–3711. 156. Bugarel,M.;Martin,A.;Fach,P.;Beutin,L.Virulencegeneprofilingofenterohemorrhagic(EHEC)and enteropathogenic(EPEC)Escherichiacolistrains:Abasisformolecularriskassessmentoftypicaland atypicalEPECstrains.BMCMicrobiol.2011,11,142. 157. Baines,D.;Sumarah,M.;Kuldau,G.;Juba,J.;Mazza,A.;Masson,L.Aflatoxin,fumonisinandShigatoxinproducingEscherichiacoliinfectionsincalvesandtheeffectivenessofcelmanax®/dairyman’schoiceTM applicationstoeliminatemorbidityandmortalitylosses.Toxins(Basel)2013,5,1872–1895. 158. Bugarel,M.;Beutin,L.;Fach,P.Low‐densitymacroarraytargetingnon‐locusofenterocyteeffacement effectors(nlegenes)andmajorvirulencefactorsofShigatoxin‐producingEscherichiacoli(STEC):Anew approachformolecularriskassessmentofSTECisolates.Appl.Environ.Microbiol.2010,76,203–211. 159. Wickham,M.E.;Lupp,C.;Mascarenhas,M.;Vázquez,A.;Coombes,B.K.;Brown,N.F.;Coburn,B.A.;Deng, W.;Puente,J.L.;Karmali,M.A.;etal.Bacterialgeneticdeterminantsofnon‐O157STECoutbreaksand hemolytic‐uremicsyndromeafterinfection.J.Infect.Dis.2006,194,819–827. 160. Buvens,G.;Piérard,D.Virulenceprofilinganddiseaseassociationofverocytotoxin‐producingEscherichia coliO157andnon‐O157isolatesinBelgium.FoodbornePathog.Dis.2012,9,530–535. 161. deBoer,R.F.;Ferdous,M.;Ott,A.;Scheper,H.R.;Wisselink,G.J.;Heck,M.E.;Rossen,J.W.;Kooistra‐Smid, A.M.D.AssessingthepublichealthriskofShigatoxin‐producingEscherichiacolibyuseofarapiddiagnostic screeningalgorithm.J.Clin.Microbiol.2015,53,1588–1598. 162. Ferdous,M.;Friedrich,A.W.;Grundmann,H.;deBoer,R.F.;Croughs,P.D.;Islam,M.A.;Kluytmans‐van denBergh,M.F.Q.;Kooistra‐Smid,A.M.D.;Rossen,J.W.A.Molecularcharacterizationandphylogenyof Shigatoxin–producingEscherichiacoliisolatesobtainedfromtwoDutchregionsusingwholegenome sequencing.Clin.Microbiol.Infect.2016,22,642.e1–642.e9. 163. Matussek,A.;Jernberg,C.;Einemo,I.‐M.;Monecke,S.;Ehricht,R.;Engelmann,I.;Löfgren,S.;Mernelius,S. GeneticmakeupofShigatoxin‐producingEscherichiacoliinrelationtoclinicalsymptomsanddurationof shedding:amicroarrayanalysisofisolatesfromSwedishchildren.Eur.J.Clin.Microbiol.Infect.Dis.2017, 36,1433–1441. 164. Vieira,M.A.;DosSantos,L.F.;Dias,R.C.B.;Camargo,C.H.;Pinheiro,S.R.S.;Gomes,T.A.T.;Hernandes,R.T. AtypicalenteropathogenicEscherichiacoliasaetiologicagentsofsporadicandoutbreak‐associated diarrhoeainBrazil.J.Med.Microbiol.2016,65,998–1006. 165. Xu,Y.;Bai,X.;Jin,Y.;Hu,B.;Wang,H.;Sun,H.;Fan,R.;Fu,S.;Xiong,Y.Highprevalenceofvirulence genesinspecificgenotypesofatypicalenteropathogenicEscherichiacoli.Front.Cell.Infect.Microbiol.2017, 7,109. 166. Buckner,M.M.C.;Croxen,M.a;Arena,E.T.;Finlay,B.B.Acomprehensivestudyofthecontributionof SalmonellaentericaserovarTyphimuriumSPI2effectorstobacterialcolonization,survival,andreplication intyphoidfever,macrophage,andepithelialcellinfectionmodels.Virulence2011,2,208–216. 167. Yang,Y.;Yu,C.;Ding,K.;Zhang,C.;Liao,C.;Jia,Y.;Li,J.;Cheng,X.RoleofthesseK1geneinthe pathogenicityofSalmonellaentericaserovarEnteritidisinvitroandinvivo.Microb.Pathog.2018,117,270– 275. 168. Zhang,X.;He,L.;Zhang,C.;Yu,C.;Yang,Y.;Jia,Y.;Cheng,X.;Li,Y.;Liao,C.;Li,J.;etal.Theimpactof sseK2deletiononSalmonellaentericaserovarTyphimuriumvirulenceinvivoandinvitro.BMCMicrobiol. 2019,19,182. 169. Lawley,T.D.;Chan,K.;Thompson,L.J.;Kim,C.C.;Govoni,G.R.;Monack,D.M.Genome‐widescreenfor Salmonellagenesrequiredforlong‐termsystemicinfectionofthemouse.PLoSPathog.2006,2,e11.
Microorganisms2020,8,35723of23 170. Nadler,C.;Baruch,K.;Kobi,S.;Mills,E.;Haviv,G.;Farago,M.;Alkalay,I.;Bartfeld,S.;Meyer,T.F.;Ben‐ Neriah,Y.;etal.ThetypeIIIsecretioneffectorNleEinhibitsNF‐κBactivation.PLoSPathog.2010,6, e1000743. 171. Ruchaud‐Sparagano,M.‐H.;Mühlen,S.;Dean,P.;Kenny,B.TheenteropathogenicE.coli(EPEC)Tireffector inhibitsNF‐κBactivitybytargetingTNFαreceptor‐associatedfactors.PLoSPathog.2011,7,e1002414. 172. Molloy,S.Bacterialpathogenesis:AsweetinteractionwithdeathforEPEC.Nat.Rev.Microbiol.2013,11, 659. 173. Pearson,J.S.;Hartland,E.L.AsurprisingsweetenerfromenteropathogenicEscherichiacoli.GutMicrobes 2014,5,766–769. 174. Scott,N.E.;Giogha,C.;Pollock,G.L.;Kennedy,C.L.;Webb,A.I.;Williamson,N.A.;Pearson,J.S.;Hartland, E.L.ThebacterialarginineglycosyltransferaseeffectorNleBpreferentiallymodifiesFas‐associateddeath domainprotein(FADD).J.Biol.Chem.2017,292,17337–17350. 175. ElQaidi,S.;Zhu,C.;McDonald,P.;Roy,A.;Maity,P.K.;Rane,D.;Perera,C.;Hardwidge,P.R.Highthroughputscreeningforbacterialglycosyltransferaseinhibitors.Front.Cell.Infect.Microbiol.2018,8,435. 176. Xu,C.;Liu,X.;Zha,H.;Fan,S.;Zhang,D.;Li,S.;Xiao,W.Apathogen‐derivedeffectormodulateshost glucosemetabolismbyarginineGlcNAcylationofHIF‐1αprotein.PLoSPathog.2018,14,e1007259. 177. Blasche,S.;Arens,S.;Ceol,A.;Siszler,G.;Schmid,M.A.;Häuser,R.;Schwarz,F.;Wuchty,S.;Aloy,P.;Uetz, P.;etal.TheEHEC‐hostinteractomerevealsnoveltargetsforthetranslocatedintiminreceptor.Sci.Rep. 2014,4,7531. 178. Law,R.J.;Law,H.T.;Scurll,J.M.;Scholz,R.;Santos,A.S.;Shames,S.R.;Deng,W.;Croxen,M.A.;Li,Y.;de Hoog,C.L.;etal.Quantitativemassspectrometryidentifiesnovelhostbindingpartnersforpathogenic EscherichiacolitypeIIIsecretionsystemeffectors.J.ProteomeRes.2016,15,1613–1622. 179. ElQaidi,S.;Scott,N.E.;Hays,M.P.;Geisbrecht,B.V.;Watkins,S.;Hardwidge,P.R.Anintra‐bacterial activityforaT3SSeffector.Sci.Rep.2020,10,1073. ©2020bytheauthors.LicenseeMDPI,Basel,Switzerland.Thisarticleisanopenaccess articledistributedunderthetermsandconditionsoftheCreativeCommonsAttribution (CCBY)license(http://creativecommons.org/licenses/by/4.0/).