Featured Publications
Pathogenic RAB34 variants impair primary cilium assembly and cause a novel oral-facial-digital syndrome
Bruel A, Ganga A, Nosková L, Valenzuela I, Martinovic J, Duffourd Y, Zikánová M, Majer F, Kmoch S, Mohler M, Sun J, Sweeney L, Martínez-Gil N, Thauvin-Robinet C, Breslow D. Pathogenic RAB34 variants impair primary cilium assembly and cause a novel oral-facial-digital syndrome. Human Molecular Genetics 2023, 32: 2822-2831. PMID: 37384395, PMCID: PMC10481091, DOI: 10.1093/hmg/ddad109.Peer-Reviewed Original ResearchConceptsCilia assemblyCiliary membrane formationIntracellular ciliogenesis pathwayPrimary cilia assemblyBi-allelic missense variantsRab proteinsRab GTPaseCiliary proteinsSmall GTPaseNascent ciliaMother centriolePrimary ciliaC-terminusProtein productsPathogenic variantsRab34Cell typesFunctional impactMissense variantsGTPaseStrong lossCiliogenesisSignificant defectsGenesKey mediatorRab34 GTPase mediates ciliary membrane formation in the intracellular ciliogenesis pathway
Ganga AK, Kennedy MC, Oguchi ME, Gray S, Oliver KE, Knight TA, De La Cruz EM, Homma Y, Fukuda M, Breslow DK. Rab34 GTPase mediates ciliary membrane formation in the intracellular ciliogenesis pathway. Current Biology 2021, 31: 2895-2905.e7. PMID: 33989527, PMCID: PMC8282722, DOI: 10.1016/j.cub.2021.04.075.Peer-Reviewed Original ResearchConceptsIntracellular pathwaysCiliary membrane biogenesisCiliary membrane formationIntracellular ciliogenesis pathwayMDCK cellsPolarized MDCK cellsDistinct molecular requirementsPrimary cilia formExtracellular pathwaysTissue-specific mannerCiliary pocketGTPase domainMembrane biogenesisDistinct functional propertiesCiliary vesiclesAssembly intermediatesCilia formSignal transductionGTP bindingMother centriolePrimary ciliaCiliogenesisDivergent residuesIntracellular ciliaRab34Mechanism and Regulation of Centriole and Cilium Biogenesis
Breslow DK, Holland AJ. Mechanism and Regulation of Centriole and Cilium Biogenesis. Annual Review Of Biochemistry 2019, 88: 1-34. PMID: 30601682, PMCID: PMC6588485, DOI: 10.1146/annurev-biochem-013118-111153.ChaptersConceptsInterphase microtubule cytoskeletonMicrotubule-based organellesBiogenesis of centriolesMost animal cellsCore of centrosomesFormation of ciliaNine-fold symmetryCilia biologyCilia biogenesisCellular signalingMicrotubule cytoskeletonAnimal cellsMitotic spindleBasal bodiesHuman diseasesCentriolesBiogenesisRegulatory controlCentral roleCiliaExciting avenuesCentrosomesCytoskeletonOrganellesSignalingA CRISPR-based screen for Hedgehog signaling provides insights into ciliary function and ciliopathies
Breslow DK, Hoogendoorn S, Kopp AR, Morgens DW, Vu BK, Kennedy MC, Han K, Li A, Hess GT, Bassik MC, Chen JK, Nachury MV. A CRISPR-based screen for Hedgehog signaling provides insights into ciliary function and ciliopathies. Nature Genetics 2018, 50: 460-471. PMID: 29459677, PMCID: PMC5862771, DOI: 10.1038/s41588-018-0054-7.Peer-Reviewed Original ResearchConceptsFunctional genomic screensGenome-wide CRISPRCiliary functionHedgehog-responsive cellsCiliary signalingΕ-tubulinProtein complexesGenomic screenEmbryonic developmentGene disruptionPrimary ciliaΔ-tubulinNovel componentCiliopathiesCRISPRCiliary structureUnbiased toolHedgehogUnifying causeScreenGenesSignalingCiliaSystematic analysisPathwayAn in vitro assay for entry into cilia reveals unique properties of the soluble diffusion barrier
Breslow DK, Koslover EF, Seydel F, Spakowitz AJ, Nachury MV. An in vitro assay for entry into cilia reveals unique properties of the soluble diffusion barrier. Journal Of Cell Biology 2013, 203: 129-147. PMID: 24100294, PMCID: PMC3798247, DOI: 10.1083/jcb.201212024.Peer-Reviewed Original ResearchMeSH KeywordsActin CytoskeletonAnimalsCell LineCell MembraneCell Membrane PermeabilityCiliaDiffusionMiceMicroscopy, FluorescenceMicroscopy, VideoModels, BiologicalMolecular WeightNuclear PoreProtein TransportProteinsRecombinant Fusion ProteinsReproducibility of ResultsTime FactorsTime-Lapse ImagingTransfectionConceptsNuclear pore complexCiliary diffusion barrierPore complexActin cytoskeletonMembrane proteinsActive transportPrimary ciliaPlasma membraneCiliary membraneSpecific proteinsLarge proteinsMechanistic basisPermeabilized cellsProteinCiliaAxon initial segmentMembraneCellsCytoskeletonInitial segmentEntryTransportAssaysVivoComplexesOrm family proteins mediate sphingolipid homeostasis
Breslow DK, Collins SR, Bodenmiller B, Aebersold R, Simons K, Shevchenko A, Ejsing CS, Weissman JS. Orm family proteins mediate sphingolipid homeostasis. Nature 2010, 463: 1048-1053. PMID: 20182505, PMCID: PMC2877384, DOI: 10.1038/nature08787.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAsthmaCell LineConserved SequenceFatty Acids, MonounsaturatedHeLa CellsHomeostasisHumansMolecular Sequence DataMultigene FamilyMultiprotein ComplexesPhosphoric Monoester HydrolasesPhosphorylationProtein BindingSaccharomyces cerevisiaeSaccharomyces cerevisiae ProteinsSerine C-PalmitoyltransferaseSphingolipidsConceptsOrm proteinsSphingolipid homeostasisSphingolipid productionFunctional genomics approachSphingolipid metabolismGenomic approachesGene familyPhosphorylation sitesORM geneORMDL genesRate-limiting enzymeRegulatory pathwaysNegative regulatorGene expressionSphingolipid synthesisSerine palmitoyltransferaseEssential roleProteinCritical mediatorGenesHomeostasisStructural componentsMetabolismMisregulationSaccharomycesA disease-associated PPP2R3C-MAP3K1 phospho-regulatory module controls centrosome function
Ganga A, Sweeney L, Rubio Ramos A, Wrinn C, Bishop C, Hamel V, Guichard P, Breslow D. A disease-associated PPP2R3C-MAP3K1 phospho-regulatory module controls centrosome function. Current Biology 2024, 34: 4824-4834.e6. PMID: 39317195, PMCID: PMC11496028, DOI: 10.1016/j.cub.2024.08.058.Peer-Reviewed Original ResearchCentrosome functionKinase-phosphatase pairSystems genetics approachDisorders of gonadal developmentCentriolar localizationCentriole proteinsGrowth defectSystems geneticsPhosphatase subunitFunctional partnersCentrosomal proteinsGene functionMicrotubule organizationCentrosome regulationGenetic approachesPPP2R3CJNK signalingCell signalingKinase activityCentrosome biogenesisAcute overexpressionGonadal developmentRegulatory mechanismsMAP3K1Gonadal dysgenesis
2015
Chapter 11 Analysis of soluble protein entry into primary cilia using semipermeabilized cells
Breslow DK, Nachury MV. Chapter 11 Analysis of soluble protein entry into primary cilia using semipermeabilized cells. Methods In Cell Biology 2015, 127: 203-221. PMID: 25837393, PMCID: PMC4797650, DOI: 10.1016/bs.mcb.2014.12.006.BooksConceptsSemipermeabilized cellsProtein entriesPrimary ciliaCiliary diffusion barrierNuclear pore complexPrimary cilia functionPore complexMammalian cellsSignal transductionSpecialized compartmentsCilia functionPlasma membraneCiliary membraneIntact cellsExperimental perturbationsCell surfaceProtein exchangeCiliaVitro systemAxon initial segmentMechanistic analysisChapter 11 AnalysisUnique resourceCapture assayCells
2014
The Intraflagellar Transport Protein IFT27 Promotes BBSome Exit from Cilia through the GTPase ARL6/BBS3
Liew GM, Ye F, Nager AR, Murphy JP, Lee JS, Aguiar M, Breslow DK, Gygi SP, Nachury MV. The Intraflagellar Transport Protein IFT27 Promotes BBSome Exit from Cilia through the GTPase ARL6/BBS3. Developmental Cell 2014, 31: 265-278. PMID: 25443296, PMCID: PMC4255629, DOI: 10.1016/j.devcel.2014.09.004.Peer-Reviewed Original ResearchConceptsCargo entryBardet-Biedl syndrome proteinsIntraflagellar transport (IFT) machineryBiochemical reconstitution assaysNucleotide-free formCiliary exitTransport machinerySyndrome proteinGTP loadingUnbiased proteomicsCoat assemblyReconstitution assaysBBSomeARL6IFT27CiliaBBS3AssemblyProteomicsMachineryProteinCargoSortingExitActivation
2013
Sphingolipid Homeostasis in the Endoplasmic Reticulum and Beyond
Breslow DK. Sphingolipid Homeostasis in the Endoplasmic Reticulum and Beyond. Cold Spring Harbor Perspectives In Biology 2013, 5: a013326. PMID: 23545423, PMCID: PMC3683901, DOI: 10.1101/cshperspect.a013326.BooksConceptsSphingolipid homeostasisEndoplasmic reticulumEssential cellular rolesSphingolipid metabolismCritical regulatory sitePotent signaling moleculesCellular rolesFamily proteinsSphingolipid productionSignaling moleculesRegulatory sitesPhysiologic cuesBasic biochemistryComplex glycosphingolipidsMembrane functionHomeostasisDiverse groupSphingolipidsNew insightsReticulumMetabolic demandsDetailed understandingMetabolismStructural componentsInitial synthesisSingle molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors
Ye F, Breslow DK, Koslover EF, Spakowitz AJ, Nelson WJ, Nachury MV. Single molecule imaging reveals a major role for diffusion in the exploration of ciliary space by signaling receptors. ELife 2013, 2: e00654. PMID: 23930224, PMCID: PMC3736543, DOI: 10.7554/elife.00654.Peer-Reviewed Original ResearchConceptsCiliary membrane proteinsIntraflagellar transportIFT trainsMembrane proteinsMembrane protein diffusionSingle-molecule imagingSomatostatin receptor 3Active transportPrimary ciliaCiliary membraneMolecule imagingProtein diffusionDynamic organizationDirectional movementReceptor 3ProteinCiliaPossible roleStatistical subtractionSingle moleculesMajor roleSmoothenedSMOSSTR3Diffusive behavior
2011
Protein kinase Ypk1 phosphorylates regulatory proteins Orm1 and Orm2 to control sphingolipid homeostasis in Saccharomyces cerevisiae
Roelants FM, Breslow DK, Muir A, Weissman JS, Thorner J. Protein kinase Ypk1 phosphorylates regulatory proteins Orm1 and Orm2 to control sphingolipid homeostasis in Saccharomyces cerevisiae. Proceedings Of The National Academy Of Sciences Of The United States Of America 2011, 108: 19222-19227. PMID: 22080611, PMCID: PMC3228448, DOI: 10.1073/pnas.1116948108.Peer-Reviewed Original ResearchConceptsProtein kinase Ypk1Orm phosphorylationSphingolipid depletionSphingolipid biosynthesisTORC2-dependent phosphorylationTemperature-sensitive growthIntegral membrane proteinsWild-type levelsOnly weak activationPrimary physiological roleMutant displaysYpk1Regulatory modificationFamily proteinsMembrane proteinsORM geneKey homeostatic regulatorsProtein kinaseSphingolipid homeostasisFirst enzymeSphingolipid productionSphingolipid synthesisEndoplasmic reticulumPhysiological rolePhosphorylationA Novel Protein LZTFL1 Regulates Ciliary Trafficking of the BBSome and Smoothened
Seo S, Zhang Q, Bugge K, Breslow DK, Searby CC, Nachury MV, Sheffield VC. A Novel Protein LZTFL1 Regulates Ciliary Trafficking of the BBSome and Smoothened. PLOS Genetics 2011, 7: e1002358. PMID: 22072986, PMCID: PMC3207910, DOI: 10.1371/journal.pgen.1002358.Peer-Reviewed Original ResearchConceptsCiliary traffickingBBS proteinsBardet-Biedl syndrome proteinsLeucine zipper transcriptionHedgehog signal transducerG protein-coupled receptorsProtein-coupled receptorsCiliary entryBBSome subunitsProtein traffickingSyndrome proteinProtein complexesCellular processesNovel proteinPrimary ciliaHedgehog signalingSignal transducerBBSomeImportant regulatorTraffickingLZTFL1Ciliary functionProteinBBS3CiliaPrimary Cilia: How to Keep the Riff-Raff in the Plasma Membrane
Breslow DK, Nachury MV. Primary Cilia: How to Keep the Riff-Raff in the Plasma Membrane. Current Biology 2011, 21: r434-r436. PMID: 21640903, DOI: 10.1016/j.cub.2011.04.039.Commentaries, Editorials and Letters
2010
Membranes in Balance: Mechanisms of Sphingolipid Homeostasis
Breslow DK, Weissman JS. Membranes in Balance: Mechanisms of Sphingolipid Homeostasis. Molecular Cell 2010, 40: 267-279. PMID: 20965421, PMCID: PMC2987644, DOI: 10.1016/j.molcel.2010.10.005.Peer-Reviewed Reviews, Practice Guidelines, Standards, and Consensus StatementsConceptsSphingolipid homeostasisCell biologyEukaryotic cell biologyKey cellular rolesComplex membrane compositionCellular rolesSecretory pathwaySphingolipid biosynthesisEnzymatic machineryPhysiologic cuesSphingolipid metabolismMembrane compositionSphingolipidsBiologyNew insightsHomeostasisStructural componentsMembraneCellsBiosynthesisDefining featureMachineryGlycerolipidsEnzymeImproved understanding
2008
A comprehensive strategy enabling high-resolution functional analysis of the yeast genome
Breslow DK, Cameron DM, Collins SR, Schuldiner M, Stewart-Ornstein J, Newman HW, Braun S, Madhani HD, Krogan NJ, Weissman JS. A comprehensive strategy enabling high-resolution functional analysis of the yeast genome. Nature Methods 2008, 5: 711-718. PMID: 18622397, PMCID: PMC2756093, DOI: 10.1038/nmeth.1234.Peer-Reviewed Original ResearchConceptsGenetic interaction studiesEssential yeast genesChemical genetic screenHigh-resolution functional analysisGenetic screenYeast genesYeast genomeHypomorphic alleleSaccharomyces cerevisiaeFunctional analysisStrain collectionGrowth competitionInteraction studiesGenomeCerevisiaeGenesAllelesScreen
2006
Single-cell proteomic analysis of S. cerevisiae reveals the architecture of biological noise
Newman JR, Ghaemmaghami S, Ihmels J, Breslow DK, Noble M, DeRisi JL, Weissman JS. Single-cell proteomic analysis of S. cerevisiae reveals the architecture of biological noise. Nature 2006, 441: 840-846. PMID: 16699522, DOI: 10.1038/nature04785.Peer-Reviewed Original ResearchConceptsSingle-cell proteomic analysisBiological noiseSingle-cell resolutionProtein abundance measurementsDNA microarray analysisProtein noise levelsSingle-cell dataHigh-throughput flow cytometryProtein-specific differencesProteomic analysisS. cerevisiaeMinimal mediumProtein abundanceMicroarray analysisCellular behaviorCellular responsesProtein synthesisMessenger RNAEnvironmental changesProtein modesProtein levelsProtein expressionFlow cytometryDetailed viewRemarkable structure