Hampel A, Tritz R, Hicks M, Cruz P
Department of Biological Sciences, Northern Illinois University, DeKalb 60115.
Nucleic Acids Res. 1990 Jan 25;18(2):299-304. doi: 10.1093/nar/18.2.299.
We have identified the catalytic domain within the sequence of the negative strand of the satellite RNA of tobacco ringspot virus. Minimum energy RNA folding calculations predict a two dimensional model with four major helical regions which are supported by mutagenesis experiments. This model for the catalytic complex consists of a 50 base catalytic RNA and a 14 base substrate RNA folded together in a type of hairpin two dimensional structure. Part of the recognition region between the catalyst and substrate is two helices of 6 bases and 4 bases respectively. Catalytic activity remains when the bases in these two helices are changed but base pairing is maintained. Thus an appropriately engineered 'hairpin' catalyst is capable of cleaving heterologous RNA.
我们已经在烟草环斑病毒卫星RNA负链序列中鉴定出催化结构域。最小能量RNA折叠计算预测出一个具有四个主要螺旋区域的二维模型,该模型得到了诱变实验的支持。这种催化复合物模型由一个50个碱基的催化RNA和一个14个碱基的底物RNA组成,它们以一种发夹二维结构折叠在一起。催化剂与底物之间识别区域的一部分分别是两个由6个碱基和4个碱基组成的螺旋。当这两个螺旋中的碱基发生变化但碱基配对得以维持时,催化活性仍然存在。因此,一个经过适当设计的“发夹”催化剂能够切割异源RNA。