B-CyclinCDKs Regulate Mitotic Spindle Assembly by Phosphorylating Kinesins-5 in Budding Yeast 英文参考文献.docVIP

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B-CyclinCDKs Regulate Mitotic Spindle Assembly by Phosphorylating Kinesins-5 in Budding Yeast 英文参考文献.doc

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B-CyclinCDKs Regulate Mitotic Spindle Assembly by Phosphorylating Kinesins-5 in Budding Yeast 英文参考文献

B-Cyclin/CDKsRegulateMitoticSpindleAssemblyby PhosphorylatingKinesins-5inBuddingYeast MarkK.Chee,StevenB.Haase* DepartmentofBiology,DukeUniversity,Durham,NorthCarolina,UnitedStatesofAmerica Abstract AlthoughithasbeenknownformanyyearsthatB-cyclin/CDKcomplexesregulatetheassemblyofthemitoticspindleand entryintomitosis,thefullcomplementofrelevantCDKtargetshasnotbeenidentified.Ithaspreviouslybeenshownina varietyofmodelsystemsthatB-typecyclin/CDKcomplexes,kinesin-5motors,andtheSCFCdc4ubiquitinligasearerequired fortheseparationofspindlepolesandassemblyofabipolarspindle.Ithasbeensuggestedthat,inbuddingyeast,B-type cyclin/CDK(Clb/Cdc28)complexespromotespindlepoleseparationbyinhibitingthedegradationofthekinesins-5Kip1and Cin8bytheanaphase-promotingcomplex(APCCdh1).Wehavedetermined,however,thattheKip1andCin8proteinsare presentatwild-typelevelsintheabsenceofClb/Cdc28kinaseactivity.Here,weshowthatKip1andCin8areinvitrotargets of Clb2/Cdc28 and that the mutation of conserved CDK phosphorylation sites on Kip1 inhibits spindle pole separation withoutaffectingtheprotein’sinvivolocalizationorabundance.MassspectrometryanalysisconfirmsthattwoCDKsitesin thetaildomainofKip1arephosphorylatedinvivo.Inaddition,wehavedeterminedthatSic1,aClb/Cdc28-specificinhibitor, is the SCFCdc4 target that inhibits spindle pole separation in cells lacking functional Cdc4. Based on these findings, we proposethatClb/Cdc28drivesspindlepoleseparationbydirectphosphorylationofkinesin-5motors. Citation: Chee MK, Haase SB (2010) B-Cyclin/CDKs Regulate Mitotic Spindle Assembly by Phosphorylating Kinesins-5 in Budding Yeast. PLoS Genet 6(5): e1000935.doi:10.1371/journal.pgen.1000935 Editor:SueBiggins,FredHutchinsonCancerResearchCenter,UnitedStatesofAmerica ReceivedApril15,2009;AcceptedApril2,2010;PublishedMay6,2010 Copyright: ? 2010 Chee, Haase. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymediu