Dynamics of PLCγ and Src Family Kinase 1 Interactions during Nuclear Envelope Formation Revealed by FRET-FLIM 英文参考文献.docVIP

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Dynamics of PLCγ and Src Family Kinase 1 Interactions during Nuclear Envelope Formation Revealed by FRET-FLIM 英文参考文献.doc

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Dynamics of PLCγ and Src Family Kinase 1 Interactions during Nuclear Envelope Formation Revealed by FRET-FLIM 英文参考文献

DynamicsofPLCcandSrcFamilyKinase1Interactions duringNuclearEnvelopeFormationRevealedbyFRET- FLIM RichardD.Byrne1*,ChristopherApplebee1,DominicL.Poccia2,Banafshe′ Larijani1* 1Cell Biophysics Laboratory, London Research Institute, Cancer Research UK, London, United Kingdom, 2Department of Biology, Amherst College, Amherst, Massachusetts,UnitedStatesofAmerica Abstract Thenuclearenvelope(NE)breaksdownandreformsduringeachmitoticcycle.AsimilarprocesshappenstothespermNE following fertilisation. The formation of the NE in both these circumstances involves endoplasmic reticulum membranes enveloping the chromatin, but PLCc-dependent membrane fusion events are also essential. Here we demonstrate the activationofPLCcbyaSrcfamilykinase(SFK1)duringNEassembly.Weshowbytime-resolvedFRETforthefirsttimethe directinvivointeractionandtemporalregulationofPLCcandSFK1inseaurchins.Asaprerequisiteforproteinactivation, thereisarapidphosphorylationofPLCconitsY783residueinresponsetoGTPinvitro.Thisphosphorylationisdependent uponSFKactivity;thusY783phosphorylationandNEassemblyaresusceptibletoSFKinhibition.Y783phosphorylationis alsoobservedonthesurfaceofthemalepronucleus(MPN)invivoduringNEformation.Togetherthecorroborativeinvivo and in vitro data demonstrate the phosphorylation and activation of PLCc by SFK1 during NE assembly. We discuss the potentialgeneralityofsuchamechanism. Citation:ByrneRD,ApplebeeC,PocciaDL,LarijaniB(2012)DynamicsofPLCcandSrcFamilyKinase1InteractionsduringNuclearEnvelopeFormationRevealed byFRET-FLIM.PLoSONE7(7):e40669.doi:10.1371/journal.pone.0040669 Editor:EladKatz,UniversityofEdinburgh,UnitedKingdom ReceivedMarch27,2012;AcceptedJune11,2012;PublishedJuly24,2012 Copyright: ? 2012 Byrne et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:TheprojectwasfundedbyCancerResearchUKcorefundingtoL