Featured Publications
Developmental Transitions Coordinate Assembly of the Coxiella burnetii Dot/Icm Type IV Secretion System
Park D, Steiner S, Shao M, Roy CR, Liu J. Developmental Transitions Coordinate Assembly of the Coxiella burnetii Dot/Icm Type IV Secretion System. Infection And Immunity 2022, 90: e00410-22. PMID: 36190257, PMCID: PMC9584302, DOI: 10.1128/iai.00410-22.Peer-Reviewed Original ResearchConceptsSmall cell variantLarge cell variantDot/Icm type IV secretion systemCell variantIntracellular replicationActive large cell variantHost cellsIntracellular bacterial pathogenType IV secretion systemDot/Icm T4SSObligate intracellular bacterial pathogenC. burnetiiCoxiella burnetiiBiphasic developmental cycleUnique biphasic developmental cycleInfectionNew host cellsSecretion systemBacterial pathogensInfectious formHost vacuoleBurnetiiMorphological changesCellsLater stagesCover Image: Coxiella burnetii encodes an LvgA‐related protein important for intracellular replication (Cellular Microbiology 06/2021)
Steiner S, Meir A, Roy C. Cover Image: Coxiella burnetii encodes an LvgA‐related protein important for intracellular replication (Cellular Microbiology 06/2021). Cellular Microbiology 2021, 23 DOI: 10.1111/cmi.13351.Peer-Reviewed Original ResearchHost cell depletion of tryptophan by IFNγ-induced Indoleamine 2,3-dioxygenase 1 (IDO1) inhibits lysosomal replication of Coxiella burnetii
Ganesan S, Roy CR. Host cell depletion of tryptophan by IFNγ-induced Indoleamine 2,3-dioxygenase 1 (IDO1) inhibits lysosomal replication of Coxiella burnetii. PLOS Pathogens 2019, 15: e1007955. PMID: 31461509, PMCID: PMC6736304, DOI: 10.1371/journal.ppat.1007955.Peer-Reviewed Original ResearchConceptsC. burnetiiC. burnetii replicationIntracellular replicationIntracellular pathogensPro-inflammatory cytokine interferon gammaIFNγ-induced genesCell-autonomous defense mechanismAbsence of IFNγCytokine interferon-gammaMost intracellular pathogensMacrophage-like cellsKynurenine metabolitesCell depletionEffector mechanismsPathogen Coxiella burnetiiInterferon gammaIFNγTryptophan availabilityHost defenseDioxygenase 1Coxiella burnetiiTHP1 cellsRestrict replicationBacterial replicationBurnetiiAnalysis of Dot/Icm Type IVB Secretion System Subassemblies by Cryoelectron Tomography Reveals Conformational Changes Induced by DotB Binding
Park D, Chetrit D, Hu B, Roy CR, Liu J. Analysis of Dot/Icm Type IVB Secretion System Subassemblies by Cryoelectron Tomography Reveals Conformational Changes Induced by DotB Binding. MBio 2020, 11: 10.1128/mbio.03328-19. PMID: 32071271, PMCID: PMC7029142, DOI: 10.1128/mbio.03328-19.Peer-Reviewed Original ResearchConceptsType IV secretion systemSecretion systemCryoelectron tomographyInner membraneDot/Icm apparatusConformational changesDot/IcmEukaryotic host cellsBacterial inner membraneWild-type cellsHost cell membraneWhole-cell contextMultiprotein nanomachineSubtomogram analysisSophisticated nanomachinesCytoplasmic substratesProtein effectorsCell polesDNA substratesSubtomogram averagingATPase complexDNA transferHost infectionStructural basisHost cellsA unique cytoplasmic ATPase complex defines the Legionella pneumophila type IV secretion channel
Chetrit D, Hu B, Christie PJ, Roy CR, Liu J. A unique cytoplasmic ATPase complex defines the Legionella pneumophila type IV secretion channel. Nature Microbiology 2018, 3: 678-686. PMID: 29784975, PMCID: PMC5970066, DOI: 10.1038/s41564-018-0165-z.Peer-Reviewed Original ResearchConceptsCytoplasmic complexType IV secretion channelType IV secretion systemInner membrane complexTranslocation of substratesCryo-electron tomographySecretion channelCell polesCytoplasmic ATPaseSecretion systemT4SS functionHexameric assemblyMembrane complexCytoplasmic channelsDNA complexesSubstrate transferT4SSChannel activationATPaseComplexesDistinct stagesAssemblyBiogenesisATPasesFurther analysisMultiple Legionella pneumophila effector virulence phenotypes revealed through high-throughput analysis of targeted mutant libraries
Shames SR, Liu L, Havey JC, Schofield WB, Goodman AL, Roy CR. Multiple Legionella pneumophila effector virulence phenotypes revealed through high-throughput analysis of targeted mutant libraries. Proceedings Of The National Academy Of Sciences Of The United States Of America 2017, 114: e10446-e10454. PMID: 29133401, PMCID: PMC5715750, DOI: 10.1073/pnas.1708553114.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBacterial ProteinsCytotoxicity, ImmunologicDisease Models, AnimalFemaleGene Expression Regulation, BacterialHigh-Throughput Nucleotide SequencingHost-Pathogen InteractionsHumansLegionella pneumophilaLegionnaires' DiseaseLoss of Function MutationMiceMice, Inbred C57BLMutagenesis, InsertionalPhenotypeVirulenceConceptsCultured host cellsHost cellsEffector proteinsHost immune systemFunction mutationsVirulence phenotypesSevere pneumoniaInfected miceBacterial clearanceEffector mutantsLarge cohortMouse modelDot/Icm type IV secretion systemImmune systemType IV secretion systemScreen resultsEffector protein activityDifferent effector proteinsGenetic complementation studiesLegionnaires' diseaseCausative agentHost pathogenesisDifferent virulence phenotypesInfectionDiseaseThe Legionella Anti-autophagy Effector RavZ Targets the Autophagosome via PI3P- and Curvature-Sensing Motifs
Horenkamp FA, Kauffman KJ, Kohler LJ, Sherwood RK, Krueger KP, Shteyn V, Roy CR, Melia TJ, Reinisch KM. The Legionella Anti-autophagy Effector RavZ Targets the Autophagosome via PI3P- and Curvature-Sensing Motifs. Developmental Cell 2015, 34: 569-576. PMID: 26343456, PMCID: PMC4594837, DOI: 10.1016/j.devcel.2015.08.010.Peer-Reviewed Original ResearchConceptsATG8 proteinsIntracellular pathogen Legionella pneumophilaPre-autophagosomal structureAtg8/LC3 proteinsPathogen Legionella pneumophilaHigh-curvature membranesMembrane transport pathwaysCytosol of cellsEffector proteinsCatalytic domainHost cytosolRavZAutophagy proteinsLC3 proteinPathogenic microbesSubstrate affinityProteinIntermediate membraneLegionella pneumophilaAutophagosomesAutophagyCytosolTransport pathwaysInterfacial activationMembraneA Screen of Coxiella burnetii Mutants Reveals Important Roles for Dot/Icm Effectors and Host Autophagy in Vacuole Biogenesis
Newton HJ, Kohler LJ, McDonough JA, Temoche-Diaz M, Crabill E, Hartland EL, Roy CR. A Screen of Coxiella burnetii Mutants Reveals Important Roles for Dot/Icm Effectors and Host Autophagy in Vacuole Biogenesis. PLOS Pathogens 2014, 10: e1004286. PMID: 25080348, PMCID: PMC4117601, DOI: 10.1371/journal.ppat.1004286.Peer-Reviewed Original ResearchConceptsHost autophagyTransposon insertionDot/Icm effectorsDot/Icm systemAutophagosome protein LC3Intracellular growth defectPathogen-occupied vacuolesTransposon insertion mutantsVacuole biogenesisEffector proteinsInsertion mutantsModification enzymesGrowth defectArrayed libraryCentral metabolismIcm systemMutantsMolecular mechanismsVisual screenProtein LC3Host cellsIntracellular replicationGenesIntracellular pathogensRegulatory systemThe Machinery at Endoplasmic Reticulum-Plasma Membrane Contact Sites Contributes to Spatial Regulation of Multiple Legionella Effector Proteins
Hubber A, Arasaki K, Nakatsu F, Hardiman C, Lambright D, De Camilli P, Nagai H, Roy CR. The Machinery at Endoplasmic Reticulum-Plasma Membrane Contact Sites Contributes to Spatial Regulation of Multiple Legionella Effector Proteins. PLOS Pathogens 2014, 10: e1004222. PMID: 24992562, PMCID: PMC4081824, DOI: 10.1371/journal.ppat.1004222.Peer-Reviewed Original ResearchAMPylation Is Critical for Rab1 Localization to Vacuoles Containing Legionella pneumophila
Hardiman CA, Roy CR. AMPylation Is Critical for Rab1 Localization to Vacuoles Containing Legionella pneumophila. MBio 2014, 5: e01035-13. PMID: 24520063, PMCID: PMC3950522, DOI: 10.1128/mbio.01035-13.Peer-Reviewed Original ResearchConceptsLCV membraneEffector proteinsGEF activityEndoplasmic reticulumIntracellular pathogen Legionella pneumophilaLegionella effector proteinsType IV secretion systemExchange factor domainLegionella pneumophilaMembrane-bound compartmentsMembrane-bound organellesPathogen Legionella pneumophilaAccumulation of GTPHost cell functionsAMPylation activityDrrA proteinRab1 proteinRab1 recruitmentLegionella effectorsNucleotidyltransferase domainAMPylationSecretion systemPosttranslational modificationsRab1GTPasePathogen signatures activate a ubiquitination pathway that modulates the function of the metabolic checkpoint kinase mTOR
Ivanov SS, Roy CR. Pathogen signatures activate a ubiquitination pathway that modulates the function of the metabolic checkpoint kinase mTOR. Nature Immunology 2013, 14: 1219-1228. PMID: 24121838, PMCID: PMC3839319, DOI: 10.1038/ni.2740.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBase SequenceCell LineCells, CulturedCytokinesEukaryotic Initiation Factor-4EGene ExpressionHost-Pathogen InteractionsImmunoblottingLegionella pneumophilaLegionnaires' DiseaseMacrophagesMiceMolecular Sequence DataMutationProto-Oncogene Proteins c-aktReverse Transcriptase Polymerase Chain ReactionRNA InterferenceSignal TransductionTOR Serine-Threonine KinasesUbiquitinationLegionella pneumophila proteins that regulate Rab1 membrane cycling
Ingmundson A, Delprato A, Lambright DG, Roy CR. Legionella pneumophila proteins that regulate Rab1 membrane cycling. Nature 2007, 450: 365-369. PMID: 17952054, DOI: 10.1038/nature06336.Peer-Reviewed Original ResearchConceptsDrrA proteinRab1 functionMembrane cyclingGTPase-activating protein activityIntracellular pathogen Legionella pneumophilaRecruitment of Rab1Pathogen-occupied vacuolesPathogen Legionella pneumophilaDistinct biochemical reactionsL. pneumophilaLegionella pneumophilaRab1 activityRab1 activationRab proteinsEukaryotic cellsExchange factorGTP hydrolysisProtein activityRab1Membrane functionBacterial replicationProteinBiochemical reactionsPneumophilaVacuolesAnkyrin Repeat Proteins Comprise a Diverse Family of Bacterial Type IV Effectors
Pan X, Lührmann A, Satoh A, Laskowski-Arce MA, Roy CR. Ankyrin Repeat Proteins Comprise a Diverse Family of Bacterial Type IV Effectors. Science 2008, 320: 1651-1654. PMID: 18566289, PMCID: PMC2514061, DOI: 10.1126/science.1158160.Peer-Reviewed Original ResearchConceptsSecretion systemL. pneumophila-containing vacuoleIntracellular pathogen Legionella pneumophilaHost cellsDifferent bacterial proteinsType IV secretion systemMicrotubule-dependent vesicular transportEukaryotic host cellsType IV effectorsPathogen Legionella pneumophilaSpecialized secretion systemsAnkyrin Repeat ProteinsANK proteinsEukaryotic cellsHomology domainEffector proteinsEukaryotic factorsRepeat proteinsInfection of macrophagesVesicular transportBacterial proteinsLate endosomesDiverse familyProteinLegionella pneumophilaAsc and Ipaf Inflammasomes Direct Distinct Pathways for Caspase-1 Activation in Response to Legionella pneumophila▿
Case CL, Shin S, Roy CR. Asc and Ipaf Inflammasomes Direct Distinct Pathways for Caspase-1 Activation in Response to Legionella pneumophila▿. Infection And Immunity 2009, 77: 1981-1991. PMID: 19237518, PMCID: PMC2681768, DOI: 10.1128/iai.01382-08.Peer-Reviewed Original ResearchHost SNAREs mediate fusion of vacuoles containing Legionella pneumophila with vesicles exiting the endoplasmic reticulum
Arasaki K, Roy C. Host SNAREs mediate fusion of vacuoles containing Legionella pneumophila with vesicles exiting the endoplasmic reticulum. The FASEB Journal 2009, 23: 867.4-867.4. DOI: 10.1096/fasebj.23.1_supplement.867.4.Peer-Reviewed Original ResearchPlasma membrane-derived vacuoleEarly secretory vesiclesSNARE complexSNARE proteinsSecretory vesiclesPlasma membrane t-SNAREFunctional SNARE complexesPlasma membrane syntaxinsEndoplasmic reticulum-derived vesiclesLegionella pneumophilaT-SNAREsSyntaxin 4Mechanism of fusionSpecialized organellesSyntaxin 2Syntaxin 3Mammalian cellsEndoplasmic reticulumSec22bSNAP23SyntaxinBacterial uptakeLocalization studiesIntracellular pathogensBacterial replicationModulation of Rab GTPase function by a protein phosphocholine transferase
Mukherjee S, Liu X, Arasaki K, McDonough J, Galán JE, Roy CR. Modulation of Rab GTPase function by a protein phosphocholine transferase. Nature 2011, 477: 103-106. PMID: 21822290, PMCID: PMC3206611, DOI: 10.1038/nature10335.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBacterial ProteinsChlorocebus aethiopsCOS CellsDiacylglycerol CholinephosphotransferaseGuanine Nucleotide Exchange FactorsHEK293 CellsHost-Pathogen InteractionsHumansLegionella pneumophilaLegionnaires' DiseaseMass SpectrometryProtein Processing, Post-TranslationalRab GTP-Binding ProteinsThe Legionella pneumophila Effector DrrA Is Sufficient to Stimulate SNARE-Dependent Membrane Fusion
Arasaki K, Toomre DK, Roy CR. The Legionella pneumophila Effector DrrA Is Sufficient to Stimulate SNARE-Dependent Membrane Fusion. Cell Host & Microbe 2012, 11: 46-57. PMID: 22264512, PMCID: PMC3266541, DOI: 10.1016/j.chom.2011.11.009.Peer-Reviewed Original ResearchConceptsMembrane transport pathwaysEndoplasmic reticulumSyntaxin proteinsFusion of ERMembrane fusionSNARE-dependent membrane fusionBacterial pathogen Legionella pneumophilaPathogen-containing vacuolesSNARE protein Sec22bIntracellular bacterial pathogen Legionella pneumophilaPathogen Legionella pneumophilaFusion of vesiclesRab1 activationNoncanonical pairingTransport pathwaysRab1 GTPasePlasma membraneVesicle fusionOrganellesDrrASec22bVesiclesLegionella pneumophilaProteinVacuolesThe Legionella Effector RavZ Inhibits Host Autophagy Through Irreversible Atg8 Deconjugation
Choy A, Dancourt J, Mugo B, O’Connor T, Isberg RR, Melia TJ, Roy CR. The Legionella Effector RavZ Inhibits Host Autophagy Through Irreversible Atg8 Deconjugation. Science 2012, 338: 1072-1076. PMID: 23112293, PMCID: PMC3682818, DOI: 10.1126/science.1227026.Peer-Reviewed Original ResearchMeSH KeywordsAdaptor Proteins, Signal TransducingAutophagyAutophagy-Related Protein 7Autophagy-Related Protein 8 FamilyAutophagy-Related ProteinsBacterial ProteinsCell Culture TechniquesCysteine ProteasesGene DeletionGlycineHEK293 CellsHost-Pathogen InteractionsHumansHydrolysisLegionella pneumophilaLegionnaires' DiseaseMicrofilament ProteinsPhagosomesUbiquitin-Activating EnzymesUbiquitin-Conjugating EnzymesConceptsATG8 proteinsIntracellular pathogen Legionella pneumophilaPathogen Legionella pneumophilaAdjacent aromatic residuesCarboxyl-terminal glycine residueAutophagosome membraneEukaryotic cellsAutophagy pathwayGlycine residueAromatic residuesIntracellular pathogensRavZAutophagyProteinLegionella pneumophilaSpecific mechanismsResiduesPathogensATG3MicrobesAtg7CytosolVacuolesPathwayPneumophilaCaspase-11 stimulates rapid flagellin-independent pyroptosis in response to Legionella pneumophila
Case CL, Kohler LJ, Lima JB, Strowig T, de Zoete MR, Flavell RA, Zamboni DS, Roy CR. Caspase-11 stimulates rapid flagellin-independent pyroptosis in response to Legionella pneumophila. Proceedings Of The National Academy Of Sciences Of The United States Of America 2013, 110: 1851-1856. PMID: 23307811, PMCID: PMC3562791, DOI: 10.1073/pnas.1211521110.Peer-Reviewed Original ResearchMeSH KeywordsAdaptor Proteins, Vesicular TransportAnimalsApoptosisApoptosis Regulatory ProteinsBone Marrow CellsCalcium-Binding ProteinsCARD Signaling Adaptor ProteinsCarrier ProteinsCaspase 1CaspasesCaspases, InitiatorCells, CulturedCytokinesCytoskeletal ProteinsEnzyme ActivationFlagellinHost-Pathogen InteractionsImmunoblottingLegionella pneumophilaMacrophagesMiceMice, Inbred C57BLMice, KnockoutMutationMyeloid Differentiation Factor 88NecrosisNLR Family, Pyrin Domain-Containing 3 ProteinReceptor, Interferon alpha-betaConceptsCaspase-11 activationCaspase-11Intracellular pathogen Legionella pneumophilaType IV secretion systemDot/IcmAdapter protein TRIFFunctional type IV secretion systemPathogen Legionella pneumophilaCaspase-1 activation pathwayNAIP/NLRC4Activation pathwayLegionella pneumophilaCaspase-11-dependent pyroptosisSecretion systemSevere defectsBacterial flagellinTreatment of macrophagesType I IFN receptorHost componentsIntracellular pathogensMicrobial signaturesPathwayIFN receptorCaspase-1Alternative pathwayThe Coxiella burnetii effector EmcB is a deubiquitinase that inhibits RIG-I signaling
Duncan-Lowey J, Crabill E, Jarret A, Reed S, Roy C. The Coxiella burnetii effector EmcB is a deubiquitinase that inhibits RIG-I signaling. Proceedings Of The National Academy Of Sciences Of The United States Of America 2023, 120: e2217602120. PMID: 36893270, PMCID: PMC10089202, DOI: 10.1073/pnas.2217602120.Peer-Reviewed Original ResearchConceptsDot/Icm systemHost-adapted pathogenUbiquitin chainsI interferon productionType I interferon productionSecretion systemHost cytosolIcm systemInterferon productionProtein secretion systemBacterial secretion systemsAcid-inducible gene IHost surveillance systemImmune sensorsObligate intracellular pathogensInducible gene ICytosolic surveillance systemsProtective immune responseEffector proteinsSpecialized organellesVacuolar nicheInhibition of RIGInnate immune sensorsSurveillance systemMammalian hosts