Roles of Dynein and Dynactin in Early Endosome Dynamics Revealed Using Automated Tracking and Global Analysis 英文参考文献.docVIP

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Roles of Dynein and Dynactin in Early Endosome Dynamics Revealed Using Automated Tracking and Global Analysis 英文参考文献.doc

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Roles of Dynein and Dynactin in Early Endosome Dynamics Revealed Using Automated Tracking and Global Analysis 英文参考文献

RolesofDyneinandDynactininEarlyEndosome DynamicsRevealedUsingAutomatedTrackingand GlobalAnalysis NeftaliFlores-Rodriguez1,SalmanS.Rogers2,DavidA.Kenwright1,2,3,ThomasA.Waigh2,3,PhilipG. Woodman1*.,VictoriaJ.Allan 1,3 . * 1Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom, 2School of Physics and Astronomy, University of Manchester, Manchester, United Kingdom,3PhotonScienceInstitute,UniversityofManchester,Manchester,UnitedKingdom Abstract Microtubule-dependentmovementiscrucialforthespatialorganizationofendosomesinmosteukaryotes,butasyetthere hasbeennosystematicanalysisofhowaparticularmicrotubulemotorcontributestoearlyendosomedynamics.Herewe tracked early endosomes labeled with GFP-Rab5 on the nanometer scale, and combined this with global, first passage probability(FPP)analysistoprovideanunbiaseddescriptionofhowtheminus-endmicrotubulemotor,cytoplasmicdynein, supportsendosomemotility.Dyneincontributestoshort-rangeendosomemovement,butinparticulardrives85–98%of long, inward translocations. For these, it requires an intact dynactin complex to allow membrane-bound p150Glued to activatedynein,sincep50over-expression,whichdisruptsthedynactincomplex,inhibitsinwardmovementeventhough dyneinandp150Glued remainmembrane-bound.Longdynein-dependentmovementsoccurviaburstsatupto,8mms21 thatarelinkedbychangesinrateorpauses.Thesepeakspeedsduringrapidinwardendosomemovementarestillseen when cellular dynein levels are 50-fold reduced by RNAi knock-down of dynein heavy chain, while the number of movementsisreduced5-fold.Altogether,thesefindingsidentifyhowdyneinhelpsdefinethedynamicsofearlyendosomes. Citation: Flores-Rodriguez N, Rogers SS, Kenwright DA, Waigh TA, Woodman PG, et al. (2011) Roles of Dynein and Dynactin in Early Endosome Dynamics RevealedUsingAutomatedTrackingandGlobalAnalysis.PLoSONE6(9):e24479.doi:10.1371/journal.pone.0024479 Editor:AnnaMariaDelprato,InstitutEurope′endeChimieetBiologie,France ReceivedJune1,2011;AcceptedAugust11,2011;Published