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 stagesAnalysis 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 cellsMultiple 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 phenotypesInfectionDiseaseA 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 systemAnkyrin 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 pneumophilaModulation 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 ProteinsSyntaxin 11 Contributes to the Interferon-Inducible Restriction of Coxiella burnetii Intracellular Infection
Ganesan S, Alvarez N, Steiner S, Fowler K, Corona A, Roy C. Syntaxin 11 Contributes to the Interferon-Inducible Restriction of Coxiella burnetii Intracellular Infection. MBio 2023, 14: e03545-22. PMID: 36728431, PMCID: PMC9972978, DOI: 10.1128/mbio.03545-22.Peer-Reviewed Original ResearchConceptsC. burnetii replicationSNARE proteinsHost cellsSyntaxin-11Cell-autonomous responsesIntracellular pathogensMembrane fusion eventsLysosome-derived organellesDefense mechanismsModel bacterial pathogenMultiple cell typesEukaryotic cellsDefense pathwaysDelivery of cargoReplication of pathogensHuman proteinsFusion eventsDissemination of pathogensFusion pathwayHost proteinsIntrinsic defense mechanismsHost vesiclesHost restriction factorsStable expressionSubcellular organelles
2024
CRISPR-Cas9-based approaches for genetic analysis and epistatic interaction studies in Coxiella burnetii.
Steiner S, Roy C. CRISPR-Cas9-based approaches for genetic analysis and epistatic interaction studies in Coxiella burnetii. MSphere 2024, e0052324. PMID: 39560384, DOI: 10.1128/msphere.00523-24.Peer-Reviewed Original ResearchIntracellular replicationHost cellsDouble mutantGenetic analysisDot/Icm type IVB secretion systemType IVB secretion systemConstruction of double mutantsLoss-of-function mutantsIntracellular replication defectsLysosome-derived vacuoleIVB secretion systemObligate intracellular bacterial pathogenRandom transposon mutagenesisTransposon insertion mutantsCRISPR-Cas9-based technologiesIntracellular bacterial pathogensLoss-of-function mutationsMammalian host cellsIntroduction of premature stop codonsPremature stop codonSilence gene expressionTargeted gene disruptionBase editing approachesEffector mutantsSecretion systemThe T4bSS of Legionella features a two-step secretion pathway with an inner membrane intermediate for secretion of transmembrane effectors
Malmsheimer S, Grin I, Bohn E, Franz-Wachtel M, Macek B, Sahr T, Smollich F, Chetrit D, Meir A, Roy C, Buchrieser C, Wagner S. The T4bSS of Legionella features a two-step secretion pathway with an inner membrane intermediate for secretion of transmembrane effectors. PLOS Pathogens 2024, 20: e1012118. PMID: 39546547, PMCID: PMC11602083, DOI: 10.1371/journal.ppat.1012118.Peer-Reviewed Original ResearchConceptsEukaryotic host cellsEffector proteinsMembrane intermediatesC-terminal secretion signalHost cellsSoluble effector proteinsCytoplasmic sideBacterial inner membraneMechanism of secretionSecretion systemSecretion signalPeriplasmic loopTransmembrane effectorSecretion pathwayT4BSSInner membraneSubcellular locationIntracellular survivalMembrane targetingProtein complexesEfficient translocationBacterial cellsProteomic analysisL. pneumophilaSecretion processInSeq analysis of defined Legionella pneumophila libraries identifies a transporter-encoding gene cluster important for intracellular replication in mammalian hosts
Moss C, Roy C. InSeq analysis of defined Legionella pneumophila libraries identifies a transporter-encoding gene cluster important for intracellular replication in mammalian hosts. MBio 2024, 15: e01955-24. PMID: 39365064, PMCID: PMC11559062, DOI: 10.1128/mbio.01955-24.Peer-Reviewed Original ResearchMammalian hostsLegionella pneumophila</i>Mutant libraryIntracellular replicationDot/Icm type IV secretion systemHost cellsType IV secretion systemInfection of mammalian cellsTransposon insertion sequencingTransposon mutant libraryOpportunistic human pathogenIV secretion systemIsogenic deletion mutantsGenetic complementation studiesHost-pathogen interactionsIntracellular bacterial pathogensMultidrug efflux pumpsMammalian infection modelsATP-binding cassetteBacteria encounterDot/Icm systemSecretion systemTransposon sequencesGene clusterHost cytosol
2010
Inhibition of pathogen-induced apoptosis by a Coxiella burnetii type IV effector protein
Lührmann A, Nogueira CV, Carey KL, Roy CR. Inhibition of pathogen-induced apoptosis by a Coxiella burnetii type IV effector protein. Proceedings Of The National Academy Of Sciences Of The United States Of America 2010, 107: 18997-19001. PMID: 20944063, PMCID: PMC2973885, DOI: 10.1073/pnas.1004380107.Peer-Reviewed Original ResearchConceptsPathogen-induced apoptosisEffector proteinsMammalian hostsL. pneumophila effector proteinsType IV effector proteinsHost cellsType IV secretion systemMammalian host cellsL. pneumophilaGain of functionProtein repertoireOrganelle traffickingRepeat familyDistinct hostsSecretion systemInfection strategiesProtozoan hostsApoptosis pathwayIntracellular bacteriaIntracellular replicationVirulence determinantsProteinLegionella pneumophilaApoptosisPathogens
2001
The DotA protein from Legionella pneumophila is secreted by a novel process that requires the Dot/Icm transporter
Nagai H, Roy C. The DotA protein from Legionella pneumophila is secreted by a novel process that requires the Dot/Icm transporter. The EMBO Journal 2001, 20: 5962-5970. PMID: 11689436, PMCID: PMC125688, DOI: 10.1093/emboj/20.21.5962.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceBacterial ProteinsBiological TransportCarrier ProteinsCell MembraneCulture Media, ConditionedElectrophoresis, Polyacrylamide GelHost-Parasite InteractionsImmunoblottingLegionella pneumophilaMacromolecular SubstancesMembrane ProteinsMolecular Sequence DataOrganellesSequence Analysis, ProteinConceptsDot/icm genesDotA proteinIcm genesDot/Icm transporterPolytopic membrane proteinsDot/IcmEukaryotic host cellsN-terminal sequencingAmino acid leader peptideLegionella pneumophilaSecretion apparatusMembrane proteinsLeader peptideMembrane vesiclesProtein secretionHost cellsProteinBacterial replicationGenesTransportersPneumophilaUnique processOrganellesCulture supernatantsSecretion
2000
Identification of Icm protein complexes that play distinct roles in the biogenesis of an organelle permissive for Legionella pneumophila intracellular growth
Coers J, Kagan J, Matthews M, Nagai H, Zuckman D, Roy C. Identification of Icm protein complexes that play distinct roles in the biogenesis of an organelle permissive for Legionella pneumophila intracellular growth. Molecular Microbiology 2000, 38: 719-736. PMID: 11115108, DOI: 10.1046/j.1365-2958.2000.02176.x.Peer-Reviewed Original ResearchConceptsPhagosome traffickingTransport apparatusHost cellsProtein-protein interactionsSubset of genesGel overlay analysisDistinct phenotypic categoriesPhagocytic host cellsIcm genesTranslocation channelReplicative organelleTwo-hybridProtein complexesSpecialized organellesTransporter functionIntracellular growthMolecular levelDistinct rolesVirulence determinantsGenesPore formationBacterial pathogensBiogenesisPhenotypic categoriesLegionella pneumophilaIdentification and Subcellular Localization of the Legionella pneumophila IcmX Protein: a Factor Essential for Establishment of a Replicative Organelle in Eukaryotic Host Cells
Matthews M, Roy C. Identification and Subcellular Localization of the Legionella pneumophila IcmX Protein: a Factor Essential for Establishment of a Replicative Organelle in Eukaryotic Host Cells. Infection And Immunity 2000, 68: 3971-3982. PMID: 10858211, PMCID: PMC101675, DOI: 10.1128/iai.68.7.3971-3982.2000.Peer-Reviewed Original ResearchConceptsEukaryotic host cellsReplicative organelleGene productsHost cellsSecretion apparatusDot/Icm proteinsDot/icm genesWild-type L. pneumophilaL. pneumophila chromosomePathogen Legionella pneumophilaHost cell parasitismImmunoblot analysisMurine bone marrow-derived macrophagesL. pneumophilaConjugal transfer systemObvious orthologsBone marrow-derived macrophagesIcm genesBacterial periplasmCell parasitismMammalian macrophagesDeletion mutantsSubcellular localizationPhagosome biogenesisMarrow-derived macrophages