title: Regulation of programmed ribosomal frameshifting by co-translational refolding RNA hairpins.
abstract: RNA structures are unwound for decoding. In the process, they can pause the elongating ribosome for regulation. An example is the stimulation of -1 programmed ribosomal frameshifting, leading to 3' direction slippage of the reading-frame during elongation, by specific pseudoknot stimulators downstream of the frameshifting site. By investigating a recently identified regulatory element upstream of the SARS coronavirus (SARS-CoV) -1 frameshifting site, it is shown that a minimal functional element with hairpin forming potential is sufficient to down-regulate-1 frameshifting activity. Mutagenesis to disrupt or restore base pairs in the potential hairpin stem reveals that base-pair formation is required for-1 frameshifting attenuation in vitro and in 293T cells. The attenuation efficiency of a hairpin is determined by its stability and proximity to the frameshifting site; however, it is insensitive to E site sequence variation. Additionally, using a dual luciferase assay, it can be shown that a hairpin stimulated +1 frameshifting when placed upstream of a +1 shifty site in yeast. The investigations indicate that the hairpin is indeed a cis-acting programmed reading-frame switch modulator. This result provides insight into mechanisms governing-1 frameshifting stimulation and attenuation. Since the upstream hairpin is unwound (by a marching ribosome) before the downstream stimulator, this study's findings suggest a new mode of translational regulation that is mediated by the reformed stem of a ribosomal unwound RNA hairpin during elongation.
authors: Institute of Biochemistry, National Chung-Hsing University, Taichung, Taiwan, Republic of China.
authors: Che-Pei
authors: CP
authors: Cho
authors: Szu-Chieh
authors: SC
authors: Lin
authors: Ming-Yuan
authors: MY
authors: Chou
authors: Hsiu-Ting
authors: HT
authors: Hsu
authors: Kung-Yao
authors: KY
authors: Chang
mesh_terms: D020029:Base Pairing; D001483:Base Sequence; D018965:Frameshifting, Ribosomal; D055029:Inverted Repeat Sequences; D059372:Nucleotide Motifs; D020871:RNA Stability; D012367:RNA, Viral / Q000737:chemistry* / Q000235:genetics*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics
chemical_list: D012367:RNA, Viral
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