Inhibition of Escherichia coli RNase P by oligonucleotide directed misfolding of RNA
Inhibition of Escherichia coli RNase P by oligonucleotide directed misfolding of RNA
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DOI：
10.1261/rna.5780503
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发表时间：
2003-12-01
期刊：
影响因子：
4.5
通讯作者：
Turner, DH
中科院分区：
文献类型：
--
作者：
Childs, JL;Poole, AW;Turner, DH
寡核苷酸定向错误折叠RNA （Oligonucleotide directed misfolding of RNA, ODMiR）利用短的寡核苷酸利用RNA折叠成具有相似自由能的不同结构的能力来抑制RNA的功能。结果表明，在含有RNase P蛋白（C5蛋白）的转录混合物中，2'- o -甲基寡核苷酸m（CAGCCUACCCGG）可以在非功能结构中捕获大肠杆菌RNase P RNA （M1 RNA）。因此，在大约200 nM处，12-mer抑制RNase P 50%的前trna加工，大约需要10倍以上的12-mer来抑制含有全长、再生RNase P RNA的RNase P。在12-mer的存在下，焦碳酸二乙酯改性显示P4中位点的修饰增加，并与P4相互作用，表明对催化活性重要的大假结的结构重排。因此，即使同源蛋白促进了rna的折叠，ODMiR方法也可以应用于rna。
Oligonucleotide directed misfolding of RNA (ODMiR) uses short oligonucleotides to inhibit RNA function by exploiting the ability of RNA to-fold into different structures with similar free energies. It is shown that the 2'-O-methyl oligonucleotide, m(CAGCCUACCCGG), can trap Escherichia coli RNase P RNA (M1 RNA) in a nonfunctional structure in a transcription mixture containing RNase P protein (C5 protein). At about 200 nM, the 12-mer thus inhibits 50% of pre-tRNA processing by RNase P. Roughly 10-fold more 12-mer is required to inhibit RNase P containing full-length, renatured RNase P RNA. Diethyl pyrocarbonate modification in the presence of 12-mer reveals increased modification of sites in and interacting with P4, suggesting a structural rearrangement of a large pseudoknot important for catalytic activity. Thus, the ODMiR method can be applied to RNAs even when folding is facilitated by a cognate protein.