Regulation of Cullin RING E3 Ubiquitin Ligases by CAND1 In Vivo 英文参考文献.docVIP

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Regulation of Cullin RING E3 Ubiquitin Ligases by CAND1 In Vivo 英文参考文献.doc

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Regulation of Cullin RING E3 Ubiquitin Ligases by CAND1 In Vivo 英文参考文献

RegulationofCullinRINGE3UbiquitinLigasesbyCAND1 InVivo YeeShinChua1.,BoonKimBoh1,2.,WanpenPonyeam1,ThiloHagen1,2* 1DepartmentofBiochemistry,YongLooLinSchoolofMedicine,NationalUniversityofSingapore,Singapore,Singapore,2NUSGraduateSchoolforIntegrativeSciences andEngineering,Singapore,Singapore Abstract CullinRINGligasesaremulti-subunitcomplexesconsistingofacullinproteinwhichformsascaffoldontowhichtheRING protein Rbx1/2 and substrate receptor subunits assemble. CAND1, which binds to cullins that are not conjugated with Nedd8andnotassociatedwithsubstratereceptors,hasbeenshowntofunctionasapositiveregulatorofCullinligasesin vivo.Twomodelshavebeenproposedtoexplainthisrequirement:(i)CAND1sequesterscullinproteinsandthusprevents autoubiquitination of substrate receptors, and (ii) CAND1 is required to promote the exchange of bound substrate receptors.Usingmammaliancells,weshowthatCAND1ispredominantlycytoplasmicallylocalizedandthatcullinsarethe majorCAND1interactingproteins.However,onlysmallamountsofCAND1bindtoCul1incells,despitelowbasallevelsof Cul1neddylationandapproximatelyequalcytoplasmicendogenousproteinconcentrationsofCAND1andCul1.Compared toF-boxproteinsubstratereceptors,bindingofCAND1toCul1invivoisweak.Furthermore,preventingbindingofF-box substrate receptors to Cul1 does not increase CAND1 binding. In conclusion, our study suggests that CAND1 does not functionbysequesteringcullinsinvivotopreventsubstratereceptorautoubiquitinationandislikelytoregulatecullinRING ligaseactivityviaalternativemechanisms. Citation:ChuaYS,BohBK,PonyeamW,HagenT(2011)RegulationofCullinRINGE3UbiquitinLigasesbyCAND1InVivo.PLoSONE6(1):e16071.doi:10.1371/ journal.pone.0016071 Editor:Anja-KatrinBielinsky,UniversityofMinnesota,UnitedStatesofAmerica ReceivedSeptember17,2010;AcceptedDecember4,2010;PublishedJanuary13,2011 Copyright: ? 2011 Chua et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andrep