Dynamic Proteomics of Human Protein Level and Localization across the Cell Cycle 英文参考文献.docVIP

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Dynamic Proteomics of Human Protein Level and Localization across the Cell Cycle 英文参考文献.doc

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Dynamic Proteomics of Human Protein Level and Localization across the Cell Cycle 英文参考文献

DynamicProteomicsofHumanProteinLeveland LocalizationacrosstheCellCycle ShlomitFarkash-Amar,EranEden¤c,ArielCohen,NaamaGeva-Zatorsky¤d,LydiaCohen,RonMilo¤b AlexSigal¤a,TamarDanon,UriAlon* , DepartmentofMolecularCellBiology,WeizmannInstituteofScience,Rehovot,Israel Abstract Regulationofproteinsacrossthecellcycleisabasicprocessincellbiology.Ithasbeendifficulttostudythisgloballyin human cells due to lack of methods to accurately follow protein levels and localizations over time. Estimates based on globalmRNAmeasurementssuggestthatonlyafewpercentofhumangeneshavecell-cycledependentmRNAlevels.Here, weuseddynamicproteomicstostudythecell-cycledependenceofproteins.Weused495clonesofahumancellline,each with a different protein tagged fluorescently at its endogenous locus. Protein level and localization was quantified in individualcellsover24hofgrowthusingtime-lapsemicroscopy.Insteadofstandardchemicalormechanicalmethodsfor cellsynchronization,weemployedin-silicosynchronizationtoplaceproteinlevelsandlocalizationonatimeaxisbetween twocelldivisions.Thisnon-perturbativesynchronizationapproach,togetherwiththehighaccuracyofthemeasurements, allowed a sensitive assay of cell-cycle dependence. We further developed a computational approach that uses texture featurestoevaluatechangesinproteinlocalizations.Wefindthat40%oftheproteinsshowedcellcycledependence,of which 11% showed changes in protein level and 35% in localization. This suggests that a broader range of cell-cycle dependentproteinsexistsinhumancellsthanwaspreviouslyappreciated.Mostofthecell-cycledependentproteinsexhibit changesincellularlocalization.Suchchangescanbeausefultoolintheregulationofthecell-cyclebeingfastandefficient. Citation:Farkash-AmarS,EdenE,CohenA,Geva-ZatorskyN,CohenL,etal.(2012)DynamicProteomicsofHumanProteinLevelandLocalizationacrosstheCell Cycle.PLoSONE7(11):e48722.doi:10.1371/journal.pone.0048722 Editor:JeanPeccoud,VirginiaTech,UnitedStatesofAmerica ReceivedJuly30,2012;AcceptedSeptember27,2012;PublishedNovem