Functional Dissection of the TBK1 Molecular Network 英文参考文献.docVIP

FunctionalDissectionoftheTBK1MolecularNetwork AdrianaGoncalves.,TilmannBu¨rckstu¨mmer*.,EvelynDixit.,RuthScheicher,MariaW.Go′rna,Evren Karayel,CristinaSugar,AlexeyStukalov,TiinaBerg,RobertKralovics,MelaniePlanyavsky,KeirynL. Bennett,JacquesColinge,GiulioSuperti-Furga* CeMM-ResearchCenterforMolecularMedicine,AustrianAcademyofSciences,Vienna,Austria Abstract TANK-bindingkinase1(TBK1)andinducibleIkB-kinase(IKK-i)arecentralregulatorsoftype-Iinterferoninduction.Theyare associated with three adaptor proteins called TANK, Sintbad (or TBKBP1) and NAP1 (or TBKBP2, AZI2) whose functional relationship to TBK1 and IKK-i is poorly understood. We performed a systematic affinity purification–mass spectrometry approachtoderiveacomprehensiveTBK1/IKK-imolecularnetwork.Themostsalientfeatureofthenetworkisthemutual exclusiveinteractionoftheadaptorswiththekinases,suggestingdistinctalternativecomplexes.Immunofluorescencedata indicated that the individual adaptors reside in different subcellular locations. TANK, Sintbad and NAP1 competed for bindingofTBK1.ThebindingsiteforallthreeadaptorswasmappedtotheC-terminalcoiled-coil2regionofTBK1.Point mutants that affect binding of individual adaptors were used to reconstitute TBK1/IKK-i-deficient cells and dissect the functionalrelevanceoftheindividualkinase-adaptoredgeswithinthenetwork.Usingamicroarray-derivedgeneexpression signatureofTBK1inresponsevirusinfectionorpoly(I:C)stimulation,wefoundthatTBK1activationwasstrictlydependent ontheintegrityoftheTBK1/TANKinteraction. Citation: Goncalves A, Bu¨rckstu¨mmer T, Dixit E, Scheicher R, Go′rna MW, et al. (2011) Functional Dissection of the TBK1 Molecular Network. PLoS ONE 6(9): e23971.doi:10.1371/journal.pone.0023971 Editor:VolkerThiel,KantonalHospitalSt.Gallen,Switzerland ReceivedJune14,2011;AcceptedJuly28,2011;PublishedSeptember8,2011 Copyright: ? 2011 Goncalves et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse