RNA aptamers that specifically bind to a 16S ribosomal RNA decoding region construct.
RNA aptamers that specifically bind to a 16S ribosomal RNA decoding region construct.
复制标题
DOI：
10.1093/nar/28.15.2902
复制
发表时间：
2000-08
影响因子：
14.9
通讯作者：
J. B. Tok;J. Cho;R. Rando
中科院分区：
文献类型：
--
作者：
J. B. Tok;J. Cho;R. Rando
RNA-RNA识别是控制许多关键生物学事件的关键过程，如翻译和核酶功能。控制RNA-RNA相互作用的识别过程可以涉及互补的Watson-Crick(WC)碱基对结合，也可以涉及通过三级结构相互作用的结合。因此，确定哪些RNA-RNA结合事件可能通过体外选择过程出现是有意义的。选择16S rRNA解码区的A位点作为靶点，既是因为它具有几个不同的RNA结构基序，也是因为它是进行密码子/反密码子识别的位置，需要同时识别mRNA和tRNA。这表明，从两个不同大小的RNA文库中可以很容易地选择一个单一的RNA分子家族。与A位点16SRRNA结构最紧密的结合适体109.2-3，其共同序列限于茎环区，该区域包含三个核苷酸，与A位点16SRRNA茎环区的四个核苷酸中的三个核苷酸互补。适体茎环上的三个核苷酸的点突变都会取消其结合能力。这些研究表明，RNA适配子109.2-3通过各自的茎环与16SRRNA的简单27nT A位编码区相互作用。最有可能的相互作用模式是通过互补的WC碱基配对，通常被称为环圈-环吻基序。没有观察到与解码区其他结构基序的高亲和力结合。
RNA-RNA recognition is a critical process in controlling many key biological events, such as translation and ribozyme functions. The recognition process governing RNA-RNA interactions can involve complementary Watson-Crick (WC) base pair binding, or can involve binding through tertiary structural interaction. Hence, it is of interest to determine which of the RNA-RNA binding events might emerge through an in vitro selection process. The A-site of the 16S rRNA decoding region was chosen as the target, both because it possesses several different RNA structural motifs, and because it is the rRNA site where codon/anticodon recognition occurs requiring recognition of both mRNA and tRNA. It is shown here that a single family of RNA molecules can be readily selected from two different sizes of RNA library. The tightest binding aptamer to the A-site 16S rRNA construct, 109.2-3, has its consensus sequences confined to a stem-loop region, which contains three nucleotides complementary to three of the four nucleotides in the stem-loop region of the A-site 16S rRNA. Point mutations on each of the three nucleotides on the stem-loop of the aptamer abolish its binding capacity. These studies suggest that the RNA aptamer 109.2-3 interacts with the simple 27 nt A-site decoding region of 16S rRNA through their respective stem-loops. The most probable mode of interaction is through complementary WC base pairing, commonly referred to as a loop-loop 'kissing' motif. High affinity binding to the other structural motifs in the decoding region were not observed.