Epigenetic Silencing of Spermatocyte-Specific and Neuronal Genes by SUMO Modification of the Transcription Factor Sp3 英文参考文献.docVIP

EpigeneticSilencingofSpermatocyte-Specificand NeuronalGenesbySUMOModificationofthe TranscriptionFactorSp3 BastianStielow1.,ImmeKru¨ger1.,RolfDiezko1,FlorianFinkernagel1,NynkeGillemans2 ,John Kong-a-San2,SjaakPhilipsen2,GuntramSuske1* 1Institute of Molecular Biology and Tumor Research, Philipps-University of Marburg, Marburg, Germany, 2Department of Cell Biology, Erasmus MC, Rotterdam, The Netherlands Abstract SUMOmodificationoftranscriptionfactorsislinkedtorepressionoftranscription.ThephysiologicalsignificanceofSUMO attachment to a particular transcriptional regulator, however, is largely unknown. We have employed the ubiquitously expressed murine transcription factor Sp3 to analyze the role of SUMOylation in vivo. We generated mice and mouse embryonic fibroblasts (MEFs) carrying a subtle point mutation in the SUMO attachment sequence of Sp3 (IKEE553 D mutation).TheE553DmutationimpedesSUMOylationofSp3atK551invivo,withoutaffectingSp3proteinlevels.Expression profilingrevealedthatspermatocyte-specificgenes,suchasDmc1andDnahc8,andneuronalgenes,includingPaqr6,Rims3, and Robo3, are de-repressed in non-testicular and extra-neuronal mouse tissues and in mouse embryonic fibroblasts expressing the SUMOylation-deficient Sp3E553D mutant protein. Chromatin immunoprecipitation experiments show that transcriptionalde-repressionofthesegenesisaccompaniedbythelossofrepressiveheterochromaticmarkssuchasH3K9 and H4K20 tri-methylation and impaired recruitment of repressive chromatin-modifying enzymes. Finally, analysis of the DNA methylation state of the Dmc1, Paqr6, and Rims3 promoters by bisulfite sequencing revealed that these genes are highlymethylatedinSp3wtMEFsbutareunmethylatedinSp3E553DMEFslinkingSUMOylationofSp3totissue-specificCpG methylation. Our results establish SUMO conjugation to Sp3 as a molecular beacon for the assembly of repression machineriestomaintaintissue-specifictranscriptionalgenesilencing. Citation: StielowB,Kru¨gerI,DiezkoR,FinkernagelF,GillemansN, etal.