title: Synergetic regulation of translational reading-frame switch by ligand-responsive RNAs in mammalian cells.
abstract: Distinct translational initiation mechanisms between prokaryotes and eukaryotes limit the exploitation of prokaryotic riboswitch repertoire for regulatory RNA circuit construction in mammalian application. Here, we explored programmed ribosomal frameshifting (PRF) as the regulatory gene expression platform for engineered ligand-responsive RNA devices in higher eukaryotes. Regulation was enabled by designed ligand-dependent conformational rearrangements of the two cis-acting RNA motifs of opposite activity in -1 PRF. Particularly, RNA elements responsive to trans-acting ligands can be tailored to modify co-translational RNA refolding dynamics of a hairpin upstream of frameshifting site to achieve reversible and adjustable -1 PRF attenuating activity. Combined with a ligand-responsive stimulator, synthetic RNA devices for synergetic translational-elongation control of gene expression can be constructed. Due to the similarity between co-transcriptional RNA hairpin folding and co-translational RNA hairpin refolding, the RNA-responsive ligand repertoire provided in prokaryotic systems thus becomes accessible to gene-regulatory circuit construction for synthetic biology application in mammalian cells.
authors.affiliation: Institute of Biochemistry, National Chung-Hsing University, 250 Kuo-Kung Road, Taichung 402, Taiwan.
authors.forename: Hsiu-Ting
authors.initials: HT
authors.lastname: Hsu
authors.affiliation: Institute of Biochemistry, National Chung-Hsing University, 250 Kuo-Kung Road, Taichung 402, Taiwan.
authors.forename: Ya-Hui
authors.initials: YH
authors.lastname: Lin
authors.affiliation: Institute of Biochemistry, National Chung-Hsing University, 250 Kuo-Kung Road, Taichung 402, Taiwan kychang@dragon.nchu.edu.tw.
authors.forename: Kung-Yao
authors.initials: KY
authors.lastname: Chang
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D018965:Frameshifting, Ribosomal; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D012313:RNA / Q000378:metabolism; D059370:RNA Folding; D016601:RNA-Binding Proteins / Q000378:metabolism; D058928:Riboswitch; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D013728:Terminator Regions, Genetic; D013806:Theophylline / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; C115000:GlcT protein, Bacillus subtilis; D008024:Ligands; D016601:RNA-Binding Proteins; D058928:Riboswitch; D014157:Transcription Factors; D012313:RNA; D012435:S-Adenosylhomocysteine; D013806:Theophylline