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. 2020 Jun 22;295(31):10741–10748. doi: 10.1074/jbc.AC120.013449

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© 2020 Kelly et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc. This article is made available via the PMC Open Access Subset for unrestricted re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the COVID-19 pandemic or until permissions are revoked in writing. Upon expiration of these permissions, PMC is granted a perpetual license to make this article available via PMC and Europe PMC, consistent with existing copyright protections.

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Figure 2. — Functional characterization of the SARS-CoV and SARS-CoV-2 −1 PRF signals. A and B, analyses of silent slippery site mutants. The efficiencies of −1 PRF promoted by the WT (U UUA AAC) and silent slippery site mutant (C CUC AAC) −1 PRF signals were assayed in HEK (A) and HeLA (B). ssM denotes silent slippery site mutant. C–E, analyses of the importance of the three stems in the −1 PRF stimulating RNA pseudoknot. Silent stem 1 (St-1, C), stem 2 (St-2, D), and stem 3 (St-3, E) mutants were assayed in HEK cells. F and G, analyses of the attenuator hairpins. AH denotes constructs that included attenuator hairpin sequences. AH mutant denotes mutants harboring the silent coding attenuator hairpin sequences shown in Fig. 1 (E and F). Assays were performed using Dual-Luciferase assays as previously described (15, 16). Each data point represents a single biological replicate comprised of three technical replicates. Error bars denote S.E. n.s, not significant.