Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor Shiftless.
Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor Shiftless.
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DOI：
10.3390/v13071230
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发表时间：
2021-06-25
期刊：
影响因子：
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通讯作者：
Brierley I
中科院分区：
文献类型：
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作者：
Napthine S;Hill CH;Nugent HCM;Brierley I
干扰素刺激基因C19 orf 66的产物Shiftless（SHFL）通过下调病毒gag/pol移码信号的效率来限制人类免疫缺陷病毒复制。在这项研究中，我们证明了细菌表达的，纯化的SHFL可以降低程序性核糖体移码在体外的各种网站，包括RNA假结依赖的信号的冠状病毒IBV，SARS-CoV和SARS-CoV-2，和蛋白质依赖的刺激的心脏病毒EMCV和TMEV的效率。SHFL还降低了小鼠白血病病毒gag/pol信号的终止密码子通读效率。使用尺寸排阻色谱法，我们证实了在体外的哺乳动物核糖体的结合纯化的蛋白质。最后，通过电泳迁移率变动分析和突变分析，我们表明，表达的SHFL具有很强的RNA结合活性，这是必要的充分的活动，在抑制移码，但没有显示出明确的特异性刺激RNA结构。
The product of the interferon-stimulated gene C19orf66, Shiftless (SHFL), restricts human immunodeficiency virus replication through downregulation of the efficiency of the viral gag/pol frameshifting signal. In this study, we demonstrate that bacterially expressed, purified SHFL can decrease the efficiency of programmed ribosomal frameshifting in vitro at a variety of sites, including the RNA pseudoknot-dependent signals of the coronaviruses IBV, SARS-CoV and SARS-CoV-2, and the protein-dependent stimulators of the cardioviruses EMCV and TMEV. SHFL also reduced the efficiency of stop-codon readthrough at the murine leukemia virus gag/pol signal. Using size-exclusion chromatography, we confirm the binding of the purified protein to mammalian ribosomes in vitro. Finally, through electrophoretic mobility shift assays and mutational analysis, we show that expressed SHFL has strong RNA binding activity that is necessary for full activity in the inhibition of frameshifting, but shows no clear specificity for stimulatory RNA structures.