Liu F, Altman S
Department of Biology, Yale University, New Haven, CT 06520, USA.
Nucleic Acids Res. 1996 Jul 15;24(14):2690-6. doi: 10.1093/nar/24.14.2690.
M1 RNA, the catalytic RNA subunit of RNase P from Escherichia coli, has been covalently linked at its 3' terminus to oligonucleotides (guide sequences) that guide the enzyme to target RNAs through hybridization with the target sequences. These constructs (M1GS RNAs) have been used to determine some minimal features of model substrates. As few as 3 bp on the 3' side of the site of cleavage in a substrate complex and 1 nt on the 5' side are required for cleavage to occur. The cytosines in the 3' terminal CCA sequence of the model substrates are important for cleavage efficiency but not cleavage site selection. A purine (base-paired or not) at the 3' side of the cleavage site is important both for cleavage site selection and efficiency. M1GS RNAs provide both a simple system for characterization of the reaction governed by M1 RNA and a tool for gene therapy.
M1 RNA是来自大肠杆菌的核糖核酸酶P的催化性RNA亚基,已在其3'末端与寡核苷酸(引导序列)共价连接,这些寡核苷酸通过与靶序列杂交将该酶引导至靶RNA。这些构建体(M1GS RNA)已用于确定模型底物的一些最小特征。底物复合物中切割位点3'侧少至3个碱基对和5'侧少至1个核苷酸是切割发生所必需的。模型底物3'末端CCA序列中的胞嘧啶对切割效率很重要,但对切割位点选择不重要。切割位点3'侧的嘌呤(无论是否碱基配对)对切割位点选择和效率都很重要。M1GS RNA既为表征由M1 RNA控制的反应提供了一个简单系统,也为基因治疗提供了一种工具。