SHH1, a Homeodomain Protein Required for DNA Methylation, As Well As RDR2, RDM4, and Chromatin Remodeling Factors, Associate with RNA Polymerase IV 英文参考文献.docVIP

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SHH1, a Homeodomain Protein Required for DNA Methylation, As Well As RDR2, RDM4, and Chromatin Remodeling Factors, Associate with RNA Polymerase IV 英文参考文献.doc

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SHH1, a Homeodomain Protein Required for DNA Methylation, As Well As RDR2, RDM4, and Chromatin Remodeling Factors, Associate with RNA Polymerase IV 英文参考文献

SHH1,aHomeodomainProteinRequiredforDNA Methylation,AsWellAsRDR2,RDM4,andChromatin RemodelingFactors,AssociatewithRNAPolymeraseIV JulieA.Law1,AjayA.Vashisht2,JamesA.Wohlschlegel2*,StevenE.Jacobsen1,3* 1Department of Molecular, Cell, and Developmental Biology, University of California Los Angeles, Los Angeles, California, United States of America, 2Department of BiologicalChemistry,DavidGeffenSchoolofMedicine,UniversityofCaliforniaLosAngeles,LosAngeles,California,UnitedStatesofAmerica,3HowardHughesMedical Institute,UniversityofCaliforniaLosAngeles,LosAngeles,California,UnitedStatesofAmerica Abstract DNA methylation is an evolutionarily conserved epigenetic modification that is critical for gene silencing and the maintenance of genome integrity. In Arabidopsis thaliana, the de novo DNA methyltransferase, DOMAINS REARRANGED METHYLTRANSFERASE 2 (DRM2), is targeted to specific genomic loci by 24nt small interfering RNAs (siRNAs) through a pathwaytermedRNA–directedDNAmethylation(RdDM).BiogenesisofthetargetingsiRNAsisthoughttobeinitiatedby theactivityoftheplant-specificRNApolymeraseIV(Pol-IV).However,themechanismthroughwhichPol-IVistargetedto specific genomic loci and whether factors other than the core Pol-IV machinery are required for Pol-IV activity remain unknown. Throughthe affinity purification of NUCLEARRNA POLYMERASE D1(NRPD1), the largestsubunit ofthe Pol-IV polymerase,wefoundthatseveralpreviouslyidentifiedRdDMcomponentsco-purifywithPol-IV,namelyRNA–DEPENDENT RNA POLYMERASE 2 (RDR2), CLASSY1 (CLSY1), and RNA–DIRECTED DNA METHYLATION 4 (RDM4), suggesting that the upstreamsiRNAgeneratingportionoftheRdDMpathwaymaybemorephysicallycoupledthanpreviouslyenvisioned.A homeodomainprotein,SAWADEEHOMEODOMAINHOMOLOG1(SHH1),wasalsofoundtoco-purifywithNRPD1;andwe demonstratethatSHH1isrequiredfordenovoandmaintenanceDNAmethylation,aswellasfortheaccumulationofsiRNAs atspecificloci,confirmingitisabonafidecomponentoftheRdDMpathway. Citation:LawJA,VashishtAA,WohlschlegelJA,JacobsenSE