title: A general strategy to inhibiting viral -1 frameshifting based on upstream attenuation duplex formation.
abstract: Viral -1 programmed ribosomal frameshifting (PRF) as a potential antiviral target has attracted interest because many human viral pathogens, including human immunodeficiency virus (HIV) and coronaviruses, rely on -1 PRF for optimal propagation. Efficient eukaryotic -1 PRF requires an optimally placed stimulator structure downstream of the frameshifting site and different strategies targeting viral -1 PRF stimulators have been developed. However, accessing particular -1 PRF stimulator information represents a bottle-neck in combating the emerging epidemic viral pathogens such as Middle East respiratory syndrome coronavirus (MERS-CoV). Recently, an RNA hairpin upstream of frameshifting site was shown to act as a cis-element to attenuate -1 PRF with mechanism unknown. Here, we show that an upstream duplex formed in-trans, by annealing an antisense to its complementary mRNA sequence upstream of frameshifting site, can replace an upstream hairpin to attenuate -1 PRF efficiently. This finding indicates that the formation of a proximal upstream duplex is the main determining factor responsible for -1 PRF attenuation and provides mechanistic insight. Additionally, the antisense-mediated upstream duplex approach downregulates -1 PRF stimulated by distinct -1 PRF stimulators, including those of MERS-CoV, suggesting its general application potential as a robust means to evaluating viral -1 PRF inhibition as soon as the sequence information of an emerging human coronavirus is available.
authors.affiliation: Institute of Biochemistry, National Chung-Hsing University, 250 Kuo-Kung Road, Taichung, 402 Taiwan.
authors.forename: Hao-Teng
authors.initials: HT
authors.lastname: Hu
authors.affiliation: Institute of Biochemistry, National Chung-Hsing University, 250 Kuo-Kung Road, Taichung, 402 Taiwan.
authors.forename: Che-Pei
authors.initials: CP
authors.lastname: Cho
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: D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism; D016373:DNA, Antisense / Q000235:genetics / Q000378:metabolism; D018965:Frameshifting, Ribosomal; D015967:Gene Expression Regulation, Viral; D057809:HEK293 Cells; D006801:Humans; D055029:Inverted Repeat Sequences; D065207:Middle East Respiratory Syndrome Coronavirus / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism
chemical_list: D016373:DNA, Antisense; D012333:RNA, Messenger; D012367:RNA, Viral; D004247:DNA