DNA Demethylation and USF Regulate the Meiosis-Specific Expression of the Mouse Miwi 英文参考文献.docVIP

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DNA Demethylation and USF Regulate the Meiosis-Specific Expression of the Mouse Miwi 英文参考文献.doc

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DNA Demethylation and USF Regulate the Meiosis-Specific Expression of the Mouse Miwi 英文参考文献

DNADemethylationandUSFRegulatetheMeiosis- SpecificExpressionoftheMouseMiwi YuHou,JiaYuan,XiangZhou,XiazhouFu,HanhuaCheng*,RongjiaZhou* DepartmentofGenetics,CollegeofLifeSciences,WuhanUniversity,Wuhan,China Abstract Miwi,amemberoftheArgonautefamily,isrequiredforinitiatingspermiogenesis;however,themechanismsthatregulate the expression of the Miwi gene remain unknown. By mutation analysis and transgenic models, we identified a 303bp proximalpromoterregionofthemouseMiwigene,whichcontrolsspecificexpressionfrommidpachytenespermatocytesto roundspermatidsduringmeiosis.WecharacterizedthebindingsitesoftranscriptionfactorsNF-Y(NuclearFactorY)andUSF (UpstreamStimulatoryFactor)withinthecorepromoterandfoundthatbothfactorsspecificallybindtoandactivatethe Miwi promoter. Methylation profiling of three CpG islands within the proximal promoter reveals a markedly inverse correlation between the methylation status of the CpG islands and germ cell type–specific expression of Miwi . CpG methylationattheUSF–bindingsitewithintheE2boxinthepromoterinhibitsthebindingofUSF.TransgenicMiwi-EGFP and endogenous Miwi reveal a subcellular co-localization pattern in the germ cells of the Miwi-EGFP transgenic mouse. Furthermore,theDNAmethylationprofileoftheMiwipromoter–driventransgeneisconsistentwiththatoftheendogenous Miwipromoter,indicatingthatMiwitransgeneisepigeneticallymodifiedthroughmethylationinvivotoensureitsspatio- temporalexpression.OurfindingssuggestthatUSFcontrolsMiwiexpressionfrommidpachytenespermatocytestoround spermatids through methylation-mediated regulation. This work identifies an epigenetic regulation mechanism for the spatio-temporalexpressionofmouseMiwiduringspermatogenesis. Citation:HouY,YuanJ,ZhouX,FuX,ChengH,etal.(2012)DNADemethylationandUSFRegulatetheMeiosis-SpecificExpressionoftheMouseMiwi .PLoS Genet8(5):e1002716.doi:10.1371/journal.pgen.1002716 Editor:GregoryS.Barsh,StanfordUniversitySchoolofMedicine,UnitedStatesofAmerica ReceivedNovember28,2011;AcceptedMarch30,2012;Publi