Cell Cycle-Dependent Microtubule-Based Dynamic Transport of Cytoplasmic Dynein in Mammalian Cells 英文参考文献.docVIP

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Cell Cycle-Dependent Microtubule-Based Dynamic Transport of Cytoplasmic Dynein in Mammalian Cells 英文参考文献.doc

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Cell Cycle-Dependent Microtubule-Based Dynamic Transport of Cytoplasmic Dynein in Mammalian Cells 英文参考文献

CellCycle-DependentMicrotubule-BasedDynamic TransportofCytoplasmicDyneininMammalianCells TakuyaKobayashi,TakashiMurayama* DepartmentofPharmacology,JuntendoUniversitySchoolofMedicine,Tokyo,Japan Abstract Background:Cytoplasmicdyneincomplexisalargemulti-subunitmicrotubule(MT)-associatedmolecularmotorinvolved in various cellular functions including organelle positioning, vesicle transport and cell division. However, regulatory mechanismofthecell-cycledependentdistributionofdyneinhasnotfullybeenunderstood. Methodology/Principal Findings: Here we report live-cell imaging of cytoplasmic dynein in HeLa cells, by expressing multifunctionalgreen fluorescentprotein(mfGFP)-tagged 74-kDaintermediate chain(IC74).IC74-mfGFP wassuccessfully incorporated into functional dynein complex. In interphase, dynein moved bi-directionally along with MTs, which might carrycargossuchastransportvesicles.AsubstantialfractionofdyneinmovedtowardcellperipherytogetherwithEB1,a memberofMTplusend-tracking proteins(+TIPs),suggesting+TIPs-mediatedtransportofdynein. Inlate-interphaseand prophase,dyneinwaslocalizedatthecentrosomesandtheradialMTarray.Inprometaphaseandmetaphase,dyneinwas localizedatspindleMTswhereitfrequentlymovedfromspindlepolestowardchromosomesorcellcortex.+TIPsmaybe involvedinthetransportofspindledyneins.Possiblekinetochoreandcorticaldyneinswerealsoobserved. Conclusions and Significance: These findings suggest that cytoplasmic dynein is transported to the site of action in preparationforthefollowingcellularevents,primarilybytheMT-basedtransport.TheMT-basedtransportmayhavegreater advantage than simple diffusion of soluble dynein in rapid and efficient transport of the limited concentration of the protein. Citation: Kobayashi T, Murayama T (2009) Cell Cycle-Dependent Microtubule-Based Dynamic Transport of Cytoplasmic Dynein in Mammalian Cells. PLoS ONE4(11):e7827.doi:10.1371/journal.pone.0007827 Editor:Anja-KatrinBielinsky,UniversityofMinnesota,UnitedStatesofAmerica ReceivedJune2