Activation of HIV Transcription by the Viral Tat Protein Requires a Demethylation Step Mediated by Lysine-specific Demethylase 1 (LSD1KDM1) 英文参考文献.docVIP

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Activation of HIV Transcription by the Viral Tat Protein Requires a Demethylation Step Mediated by Lysine-specific Demethylase 1 (LSD1KDM1) 英文参考文献.doc

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Activation of HIV Transcription by the Viral Tat Protein Requires a Demethylation Step Mediated by Lysine-specific Demethylase 1 (LSD1KDM1) 英文参考文献

ActivationofHIVTranscriptionbytheViralTatProtein RequiresaDemethylationStepMediatedbyLysine- specificDemethylase1(LSD1/KDM1) NaokiSakane1,2,Hye-SookKwon1,SaraPagans1,KatrinKaehlcke1,YasuhiroMizusawa2 ,Masafumi Kamada2,KaraG.Lassen1,JonathanChan1,WarnerC.Greene1,3,4,MartinaSchnoelzer5,MelanieOtt1,3* 1Gladstone Institute of Virology and Immunology, University of California, San Francisco, California, United States of America, 2Pharmaceutical Frontier Research Laboratory, Yokohama, Japan, 3Department of Medicine, University of California, San Francisco, United States of America, 4Department of Microbiology and Immunology, University of California, San Francisco, United States of America, 5Functional Proteome Analysis, German Cancer Research Center (DKFZ), Heidelberg, Germany Abstract The essential transactivator function of the HIV Tat protein is regulated by multiple posttranslational modifications. Although individual modifications are well characterized, their crosstalk and dynamics of occurrence during the HIV transcriptioncycleremainunclear. Weexamineinteractions betweentwocriticalmodificationswithintheRNA-binding domainofTat:monomethylationoflysine51(K51)mediatedbySet7/9/KMT7,anearlyeventintheTattransactivationcycle thatstrengthenstheinteractionofTatwithTARRNA,andacetylationoflysine50(K50)mediatedbyp300/KAT3B,alater process that dissociates the complex formed by Tat, TAR RNA and the cyclin T1 subunit of the positive transcription elongationfactorb(P-TEFb).WefindK51monomethylationinhibitedinsyntheticTatpeptidescarryinganacetylgroupat K50 while acetylation can occur in methylated peptides, albeit at a reduced rate. To examine whether Tat is subject to sequentialmonomethylationandacetylationincells,weperformedmassspectrometryonimmunoprecipitatedTatproteins andgeneratednewmodification-specificTatantibodiesagainstmonomethylated/acetylatedTat.NobimodifiedTatprotein was detected in cells pointing to a demethylation step during the Tat transactivation cycle. We iden