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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Triplex structures in an RNA pseudoknot enhance mechanical stability and increase efficiency of -1 ribosomal frameshifting
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Triplex structures in an RNA pseudoknot enhance mechanical stability and increase efficiency of -1 ribosomal frameshifting

机译:RNA假结中的三链体结构可增强机械稳定性并提高-1核糖体移码的效率

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摘要

Many viruses use programmed -1 ribosomal frameshifting to express defined ratios of structural and enzymatic proteins. Pseudoknot structures in messenger RNAs stimulate frameshifting in upstream slippery sequences. The detailed molecular determinants of pseudoknot mechanical stability and frameshifting efficiency are not well understood. Here we use single-molecule unfolding studies by optical tweezers, and frameshifting assays to elucidate how mechanical stability of a pseudoknot and its frameshifting efficiency are regulated by tertiary stem-loop interactions. Mechanical unfolding of a model pseudoknot and mutants designed to dissect specific interactions reveals that mechanical stability depends strongly on triplex structures formed by stem-loop interactions. Combining single-molecule and mutational studies facilitates the identification of pseudoknot folding intermediates. Average unfolding forces of the pseudoknot and mutants ranging from 50 to 22 picoNewtons correlated with frameshifting efficiencies ranging from 53% to 0%. Formation of major-groove and minor-groove triplex structures enhances pseudoknot stem stability and torsional resistance, and may thereby stimulate frameshifting. Better understanding of the molecular determinants of frameshifting efficiency may facilitate the development of anti-virus therapeutics targeting frameshifting.
机译:许多病毒使用程序化的-1核糖体移码来表达结构蛋白和酶蛋白的确定比例。信使RNA中的假结结构刺激上游滑序列中的移码。假结机械稳定性和移码效率的详细分子决定因素尚未得到很好的理解。在这里,我们使用光学镊子进行单分子展开研究,并使用移码测定法来阐明伪结的机械稳定性及其移码效率是如何通过三级茎-环相互作用调节的。机械假结模型和旨在分析特定相互作用的突变体的机械展开显示,机械稳定性在很大程度上取决于由茎-环相互作用形成的三链体结构。将单分子和突变研究相结合有助于鉴定假结折叠中间体。假结和突变体的平均展开力范围为50到22微微牛顿,与移码效率范围为53%到0%相关。主槽和副槽三重结构的形成增强了假结茎的稳定性和抗扭强度,并可能因此刺激移码。更好地了解移码效率的分子决定因素可能有助于开发针对移码的抗病毒治疗药物。

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  • 作者单位

    Department of Chemistry, University of California, Berkeley, CA 94720;

    Graduate Institute of Biochemistry, National Chung-Hsing University, 250 Kuo-Kung Road, Taichung, 402 Taiwan;

    Graduate Institute of Biochemistry, National Chung-Hsing University, 250 Kuo-Kung Road, Taichung, 402 Taiwan;

    Department of Chemistry, University of California, Berkeley, CA 94720 Departments of Physics, Molecular and Cell Biology, and Howard Hughes Medical Institute, University of California, Berkeley, CA 94720;

    Department of Chemistry, University of California, Berkeley, CA 94720;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    optical tweezers; RNA triplexes; single-molecule; RNA folding;

    机译:光学镊子;RNA三链体;单分子RNA折叠;

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