2024
Cryo-EM structures reveal how phosphate release from Arp3 weakens actin filament branches formed by Arp2/3 complex
Chavali S, Chou S, Cao W, Pollard T, De La Cruz E, Sindelar C. Cryo-EM structures reveal how phosphate release from Arp3 weakens actin filament branches formed by Arp2/3 complex. Nature Communications 2024, 15: 2059. PMID: 38448439, PMCID: PMC10918085, DOI: 10.1038/s41467-024-46179-x.Peer-Reviewed Original ResearchMeSH KeywordsActin CytoskeletonActin-Related Protein 2-3 ComplexActinsCryoelectron MicroscopyCytoskeletonPhosphatesConceptsArp2/3 complexActin filamentsCryo-EM structureMother filamentDaughter filamentArp2/3 complex nucleates branched actin filamentsActin filament branchingBranched actin filamentsDissociation of PiADP-PiFilament branchingOrganelle movementADP stateBranch junctionsArp3A-resolutionActinArp2/3ADP-BeFxFilamentsADPPhosphate releaseFilament mechanismArp2OrganellesNoncanonical interaction with microtubules via the N-terminal nonmotor domain is critical for the functions of a bidirectional kinesin
Singh S, Siegler N, Pandey H, Yanir N, Popov M, Goldstein-Levitin A, Sadan M, Debs G, Zarivach R, Frank G, Kass I, Sindelar C, Zalk R, Gheber L. Noncanonical interaction with microtubules via the N-terminal nonmotor domain is critical for the functions of a bidirectional kinesin. Science Advances 2024, 10: eadi1367. PMID: 38324691, PMCID: PMC10849588, DOI: 10.1126/sciadv.adi1367.Peer-Reviewed Original ResearchMeSH KeywordsCryoelectron MicroscopyKinesinsMicrotubulesSaccharomyces cerevisiaeSaccharomyces cerevisiae ProteinsConceptsBidirectional motilityKinesin-5Plus-end-directed motilityKinesin-5 motorsCryo-EMC-terminal tailCryo-EM mapsPlus-end-directed kinesinCryo-electron microscopyDeletion mutantsNoncanonical interactionsIntracellular functionsCin8Cryo-electronMotor domainMotilityNonmotor domainsMutantsB-tubulinSingle-moleculeCell viabilityIn vivoIn vitroIn vitro experimentsLocal defects
2020
Dynamic and asymmetric fluctuations in the microtubule wall captured by high-resolution cryoelectron microscopy
Debs GE, Cha M, Liu X, Huehn AR, Sindelar CV. Dynamic and asymmetric fluctuations in the microtubule wall captured by high-resolution cryoelectron microscopy. Proceedings Of The National Academy Of Sciences Of The United States Of America 2020, 117: 16976-16984. PMID: 32636254, PMCID: PMC7382274, DOI: 10.1073/pnas.2001546117.Peer-Reviewed Original ResearchStructures of cofilin-induced structural changes reveal local and asymmetric perturbations of actin filaments
Huehn AR, Bibeau JP, Schramm AC, Cao W, De La Cruz EM, Sindelar CV. Structures of cofilin-induced structural changes reveal local and asymmetric perturbations of actin filaments. Proceedings Of The National Academy Of Sciences Of The United States Of America 2020, 117: 1478-1484. PMID: 31900364, PMCID: PMC6983403, DOI: 10.1073/pnas.1915987117.Peer-Reviewed Original ResearchMeSH KeywordsActin CytoskeletonActin Depolymerizing FactorsAnimalsBinding SitesCryoelectron MicroscopyHumansPhosphorylationProtein BindingRabbitsConceptsFilament severingActin filamentsSevering activityCofilin/ADF familyActin conformational changesActin filament severingFilament-severing activityCryo-electron microscopy dataSevers actin filamentsWeak severing activityUnique binding modeCofilin clustersActin structuresCofilin bindingCofilin-decorated segmentsCofilinMolecular understandingBarbed endsConformational changesCooperative bindingBinding cooperativityFilament endsPositive cooperativityBinding modesSevering
2019
Structural basis for the clamping and Ca2+ activation of SNARE-mediated fusion by synaptotagmin
Grushin K, Wang J, Coleman J, Rothman JE, Sindelar CV, Krishnakumar SS. Structural basis for the clamping and Ca2+ activation of SNARE-mediated fusion by synaptotagmin. Nature Communications 2019, 10: 2413. PMID: 31160571, PMCID: PMC6546687, DOI: 10.1038/s41467-019-10391-x.Peer-Reviewed Original ResearchConceptsCryo-electron microscopy structureActivation of SNAREsDependent membrane interactionsAnionic lipid headgroupsFusion clampActivator functionSNARE bundleSNARE proteinsMicroscopy structureC2B domainStructural basisSynaptotagmin-1SNAREpinsAliphatic loopsMembrane interactionsComplete assemblyLipid headgroupsLipid membranesNeurotransmitter releaseMembraneKey determinantSynaptotagminSyt1Calcium influxPartial insertion
2018
Structural basis of the filamin A actin-binding domain interaction with F-actin
Iwamoto DV, Huehn A, Simon B, Huet-Calderwood C, Baldassarre M, Sindelar CV, Calderwood DA. Structural basis of the filamin A actin-binding domain interaction with F-actin. Nature Structural & Molecular Biology 2018, 25: 918-927. PMID: 30224736, PMCID: PMC6173970, DOI: 10.1038/s41594-018-0128-3.Peer-Reviewed Original ResearchMeSH KeywordsActinsCryoelectron MicroscopyFilaminsHumansModels, MolecularMutation, MissenseProtein DomainsConceptsActin-binding domainCalponin homology domainHomology domainF-actinActin cross-linking proteinFunction mutationsTandem calponin homology domainsDisease-associated mutantsCryo-electron microscopyHigh-resolution structuresNumerous genetic diseasesSequence conservationHigher-order structureLinking proteinStructural basisDomain interactionsCell shapeActin filamentsMolecular understandingN-terminalFunctional studiesGenetic diseasesMissense mutationsMutationsAtomic resolutionThe actin filament twist changes abruptly at boundaries between bare and cofilin-decorated segments
Huehn A, Cao W, Elam WA, Liu X, De La Cruz EM, Sindelar CV. The actin filament twist changes abruptly at boundaries between bare and cofilin-decorated segments. Journal Of Biological Chemistry 2018, 293: 5377-5383. PMID: 29463680, PMCID: PMC5900768, DOI: 10.1074/jbc.ac118.001843.Peer-Reviewed Original ResearchMeSH KeywordsActin CytoskeletonActin Depolymerizing FactorsAnimalsCryoelectron MicroscopyProtein Structure, QuaternaryRabbitsConceptsCofilin-decorated segmentsConformational changesCofilin/ADF proteinsActin-remodeling proteinsBind actin filamentsActin filament interactionsCofilin-induced changesEffects of cofilinCooperative conformational changesProtein occupancyADF proteinsCellular processesCell divisionStructure-based methodsCryo-EMActin segmentsIntracellular transportActin filamentsFilament twistCooperative bindingCofilinTwist changesActinFluorophore labelingSubunitsHigh-resolution cryo-EM structures of actin-bound myosin states reveal the mechanism of myosin force sensing
Mentes A, Huehn A, Liu X, Zwolak A, Dominguez R, Shuman H, Ostap EM, Sindelar CV. High-resolution cryo-EM structures of actin-bound myosin states reveal the mechanism of myosin force sensing. Proceedings Of The National Academy Of Sciences Of The United States Of America 2018, 115: 1292-1297. PMID: 29358376, PMCID: PMC5819444, DOI: 10.1073/pnas.1718316115.Peer-Reviewed Original ResearchConceptsN-terminal subdomainHigh-resolution cryo-EM structuresADP stateNear-atomic resolution structuresCryo-EM structureCryo-electron microscopyHigh-resolution structuresIsoform-dependent mannerFilamentous actinResolution structureStructural basisMyosin IBActin filamentsStructural diversityRelease pathwayADP releaseActinPointed endPotent stabilizerMyosin
2017
Phosphomimetic S3D cofilin binds but only weakly severs actin filaments
Elam WA, Cao W, Kang H, Huehn A, Hocky GM, Prochniewicz E, Schramm AC, Negrón K, Garcia J, Bonello TT, Gunning PW, Thomas DD, Voth GA, Sindelar CV, De La Cruz EM. Phosphomimetic S3D cofilin binds but only weakly severs actin filaments. Journal Of Biological Chemistry 2017, 292: 19565-19579. PMID: 28939776, PMCID: PMC5712599, DOI: 10.1074/jbc.m117.808378.Peer-Reviewed Original ResearchConceptsActin bindingWild-type cofilinActin filament severingHigh cooperativitySubstitution of serineCofilin bindsActin cytoskeletonProtein cofilinCell divisionSer-3Filament severingAtom molecular dynamics simulationsSubunit interactionsN-terminusCofilinBiological processesActin filamentsTime-resolved phosphorescence anisotropyElectron cryomicroscopyRapid remodelingPhosphorylationSeveringFilament mechanical propertiesActin segmentsFilamentsStructural basis of cooperativity in kinesin revealed by 3D reconstruction of a two-head-bound state on microtubules
Liu D, Liu X, Shang Z, Sindelar CV. Structural basis of cooperativity in kinesin revealed by 3D reconstruction of a two-head-bound state on microtubules. ELife 2017, 6: e24490. PMID: 28504639, PMCID: PMC5459574, DOI: 10.7554/elife.24490.Peer-Reviewed Original Research
2014
Site-specific cation release drives actin filament severing by vertebrate cofilin
Kang H, Bradley MJ, Cao W, Zhou K, Grintsevich EE, Michelot A, Sindelar CV, Hochstrasser M, De La Cruz EM. Site-specific cation release drives actin filament severing by vertebrate cofilin. Proceedings Of The National Academy Of Sciences Of The United States Of America 2014, 111: 17821-17826. PMID: 25468977, PMCID: PMC4273407, DOI: 10.1073/pnas.1413397111.Peer-Reviewed Original ResearchMeSH KeywordsActin CytoskeletonCationsCell MovementChromatography, AffinityCofilin 1Cryoelectron MicroscopyHumansModels, MolecularSaccharomyces cerevisiaeConceptsFilament severingActin filamentsActin filament severingKey regulatory functionsConcentration of endsActin filament fragmentationEukaryotic cellsCation-binding sitesProtein cofilinDeletion mutantsS. cerevisiaeSubunit exchangeFilament turnoverActin polymerizationEssential functionsSite-specific interactionsCofilinMolecular mechanismsAssembly dynamicsRegulatory functionsActin moleculesFilament fragmentationFilament structureSustained motilitySevering
2012
Optimal noise reduction in 3D reconstructions of single particles using a volume-normalized filter
Sindelar CV, Grigorieff N. Optimal noise reduction in 3D reconstructions of single particles using a volume-normalized filter. Journal Of Structural Biology 2012, 180: 26-38. PMID: 22613568, PMCID: PMC3498508, DOI: 10.1016/j.jsb.2012.05.005.Peer-Reviewed Original ResearchConceptsParticle mapsOptimal noise reductionWiener filterError reduction schemeConventional Wiener filterNoise reductionNovel filterInversion methodNew filterHigh noise levelsParticle densityParticle volumeFilterParticlesNoise ratioNoise levelTheoretical analysisSingle particleAccurate implementationSpectral signalsSSNRInterior regionImage dataSignalsFourier inversion method
2011
An adaptation of the Wiener filter suitable for analyzing images of isolated single particles
Sindelar CV, Grigorieff N. An adaptation of the Wiener filter suitable for analyzing images of isolated single particles. Journal Of Structural Biology 2011, 176: 60-74. PMID: 21757012, PMCID: PMC3184790, DOI: 10.1016/j.jsb.2011.06.010.Peer-Reviewed Original ResearchMeSH KeywordsAlgorithmsCryoelectron MicroscopyFourier AnalysisImage Processing, Computer-AssistedModels, MolecularProtein ConformationSignal-To-Noise RatioConceptsWiener filterConventional Wiener filterCryo-EM applicationsNew filterFourier ring correlationParticle areaEstimation schemeFilterLeast-squares senseEstimation methodNoise ratioSpecific applicationsSingle particleParticlesImage fieldError reductionNumeric testsApplicationsImaging conditionsSimulated dataInput data
2010
An atomic-level mechanism for activation of the kinesin molecular motors
Sindelar CV, Downing KH. An atomic-level mechanism for activation of the kinesin molecular motors. Proceedings Of The National Academy Of Sciences Of The United States Of America 2010, 107: 4111-4116. PMID: 20160108, PMCID: PMC2840164, DOI: 10.1073/pnas.0911208107.Peer-Reviewed Original Research
2007
The beginning of kinesin's force-generating cycle visualized at 9-Å resolution
Sindelar CV, Downing KH. The beginning of kinesin's force-generating cycle visualized at 9-Å resolution. Journal Of Cell Biology 2007, 177: 377-385. PMID: 17470637, PMCID: PMC2064809, DOI: 10.1083/jcb.200612090.Peer-Reviewed Original ResearchConceptsSwitch II helixMicrotubule-binding proteinN-terminal extensionII helixNucleotide-free stateCryo-electron microscopySwitch IMicrotubule contactsResponse elementSingle-particle reconstructionConformational changesMicrotubulesActivation mechanismKinesinHigh-resolution characterizationHelixProtein