Cdc5-Dependent Asymmetric Localization of Bfa1 Fine-Tunes Timely Mitotic Exit 英文参考文献.docVIP

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Cdc5-Dependent Asymmetric Localization of Bfa1 Fine-Tunes Timely Mitotic Exit 英文参考文献.doc

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Cdc5-Dependent Asymmetric Localization of Bfa1 Fine-Tunes Timely Mitotic Exit 英文参考文献

Cdc5-DependentAsymmetricLocalizationofBfa1Fine- TunesTimelyMitoticExit JunwonKim1.,GuangmingLuo1.,YoungYilBahk2,KiwonSong1* 1DepartmentofBiochemistry,CollegeofBiotechnologyandLifeScience,YonseiUniversity,Seoul,Korea,2DepartmentofBiotechnology,KonkukUniversity,Chungju, Korea Abstract In budding yeast, the major regulator of the mitotic exit network (MEN) is Tem1, a GTPase, which is inhibited by the GTPase-activating protein (GAP), Bfa1/Bub2. Asymmetric Bfa1 localization to the bud-directed spindle pole body (SPB) duringmetaphase alsocontrols mitoticexit,butthemolecular mechanism andfunction ofthislocalization arenotwell understood, particularly in unperturbed cells. Weidentified four novel Cdc5 target residues within the Bfa1 C-terminus: 452S, 453S, 454S,and 559S.ABfa1mutantinwhichalloftheseresidueshadbeenchangedtoalanine(Bfa14A)persistedon both SPBs at anaphase and was hypo-phosphorylated, despite retaining its GAP activity for Tem1. A Bfa1 phospho- mimeticmutantinwhichalloftheseresidueswereswitchedtoaspartate(Bfa14D)alwayslocalizedasymmetricallytothe SPB. These observations demonstrate that asymmetric localization of Bfa1 is tightly linked to its Cdc5-dependent phosphorylation, but not to its GAP activity. Consistent with this, in kinase-defective cdc5-2 cells Bfa1 was not phosphorylatedandlocalizedtobothSPBs,whereasBfa14Dwasasymmetricallylocalized.BFA14Acellsprogressedthrough anaphasenormallybutdisplayeddelayedmitoticexitinunperturbedcellcycles,whileBFA14Dcellsunderwentmitoticexit withthesamekineticsaswild-typecells.WesuggestthatCdc5inducestheasymmetricdistribution ofBfa1tothebud- directed SPB independently of Bfa1 GAP activity at anaphase and that Bfa1 asymmetry fine-tunes the timing of MEN activation inunperturbed cell cycles. Citation: Kim J,Luo G,BahkYY, SongK(2012) Cdc5-DependentAsymmetric Localization ofBfa1 Fine-Tunes Timely MitoticExit. PLoSGenet 8(1): e1002450. doi:10.1371/journal.pgen.1002450 Editor:OrnaCohen-Fix,NationalInstituteofDiabetes