Sekiguchi A, Komatsu Y, Koizumi M, Ohtsuka E
Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
Nucleic Acids Res. 1991 Dec 25;19(24):6833-8. doi: 10.1093/nar/19.24.6833.
Several mutants for the minus strands of the self-cleaving domain of the satellite RNA of tobacco ringspot virus have been synthesized by joining chemically synthesized oligoribonucleotides with RNA ligase. Kinetic properties of the enzyme strands (50 nucleotides) against substrates (15-mer and 18-mer) were investigated. Structural properties of the unpaired part in the cleavage region were estimated from mutagenesis. The catalytic domain alone was proved to be responsible for the rejoining reaction of cleaved substrates. It was also found that the ribozyme could be divided into two strands without loss of activity. Effects of concentration of magnesium ion and polyamines on the cleavage reaction for the two-stranded ribozyme are also reported.
通过用RNA连接酶连接化学合成的寡核糖核苷酸，已经合成了烟草环斑病毒卫星RNA自我切割结构域负链的几种突变体。研究了酶链（50个核苷酸）对底物（15聚体和18聚体）的动力学性质。通过诱变估计了切割区域中未配对部分的结构性质。单独的催化结构域被证明对切割底物的重新连接反应负责。还发现核酶可以分成两条链而不丧失活性。还报道了镁离子和多胺浓度对双链核酶切割反应的影响。
