title: Small activating RNA binds to the genomic target site in a seed-region-dependent manner.
abstract: RNA activation (RNAa) is the upregulation of gene expression by small activating RNAs (saRNAs). In order to investigate the mechanism by which saRNAs act in RNAa, we used the progesterone receptor (PR) gene as a model, established a panel of effective saRNAs and assessed the involvement of the sense and antisense strands of saRNA in RNAa. All active saRNAs had their antisense strand effectively incorporated into Ago2, whereas such consistency did not occur for the sense strand. Using a distal hotspot for saRNA targeting at 1.6-kb upstream from the PR transcription start site, we further established that gene activation mediated by saRNA depended on the complementarity of the 5' region of the antisense strand, and that such activity was largely abolished by mutations in this region of the saRNA. We found markedly reduced RNAa effects when we created mutations in the genomic target site of saRNA PR-1611, thus providing evidence that RNAa depends on the integrity of the DNA target. We further demonstrated that this saRNA bound the target site on promoter DNA. These results demonstrated that saRNAs work via an on-site mechanism by binding to target genomic DNA in a seed-region-dependent manner, reminiscent of miRNA-like target recognition.
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, PR China.
authors.forename: Xing
authors.initials: X
authors.lastname: Meng
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, PR China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Jiang
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, PR China.
authors.forename: Nannan
authors.initials: N
authors.lastname: Chang
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, PR China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, PR China.
authors.forename: Chujun
authors.initials: C
authors.lastname: Liu
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, PR China.
authors.forename: Jingwei
authors.initials: J
authors.lastname: Xiong
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, PR China caohuiqing@pku.edu.cn.
authors.forename: Huiqing
authors.initials: H
authors.lastname: Cao
authors.affiliation: Institute of Molecular Medicine, Peking University, Beijing 100871, PR China Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, PR China liangz@pku.edu.cn.
authors.forename: Zicai
authors.initials: Z
authors.lastname: Liang
mesh_terms: D060565:Argonaute Proteins / Q000235:genetics / Q000378:metabolism; D020029:Base Pairing; D001483:Base Sequence; D001665:Binding Sites; D064113:CRISPR-Cas Systems; D002843:Chromatin / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D061986:MCF-7 Cells; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D058727:RNA, Small Untranslated / Q000235:genetics* / Q000378:metabolism; D011980:Receptors, Progesterone / Q000235:genetics* / Q000378:metabolism; D024363:Transcription Initiation Site
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; D002843:Chromatin; D058727:RNA, Small Untranslated; D011980:Receptors, Progesterone; D004247:DNA; D008156:Luciferases