大型文库揭示了针对RNA识别问题的多种解决方案。
Large libraries reveal diverse solutions to an RNA recognition problem.
作者信息
Barrick J E, Takahashi T T, Ren J, Xia T, Roberts R W
机构信息
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena CA 91125, USA.
出版信息
Proc Natl Acad Sci U S A. 2001 Oct 23;98(22):12374-8. doi: 10.1073/pnas.221467798.
Abstract
RNA loops that adopt a characteristic GNRA "tetraloop" fold are common in natural RNAs. Here, we have used in vitro selection by means of mRNA-peptide fusions to select peptides that bind an example of this RNA loop motif. Starting with the RNA recognition domain from the lambda N protein, we have constructed libraries containing 150, 1,600, and 9 trillion different peptide sequences as mRNA-peptide fusions and isolated those capable of high-affinity RNA binding. These selections have resulted in more than 80 different peptides that bind the same RNA loop. The highest affinity peptides exhibit low nanomolar dissociation constants as well as the ability to discriminate RNA hairpins differing by a single loop nucleotide. Thus, our work demonstrates that numerous, chemically distinct solutions exist for a particular RNA recognition problem.
摘要
具有特征性GNRA“四环”折叠的RNA环在天然RNA中很常见。在这里，我们通过mRNA-肽融合体外筛选，以选择与这种RNA环基序实例结合的肽。从λ N蛋白的RNA识别结构域开始，我们构建了包含150、1600和9万亿种不同肽序列的mRNA-肽融合文库，并分离出能够高亲和力结合RNA的肽。这些筛选产生了80多种与同一RNA环结合的不同肽。亲和力最高的肽表现出低纳摩尔解离常数，以及区分仅相差一个环核苷酸的RNA发夹的能力。因此，我们的工作表明，针对特定的RNA识别问题存在众多化学性质不同的解决方案。
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