Savinkova L K, Efimova L Iu, Knorre V L, Salganik R I
Mol Biol (Mosk). 1978 Nov-Dec;12(6):1313-8.
It was shown previously that E. coli RNA-polymerase being incubated with the random oligonucleotide mixtures of definite length binds certain oligoribonucleotides with the length greater than or equal to 5 nucleotides. The data presented demonstrate that T7 phage induced RNA-polymerase (T7 RNA-polymerase) also binds selectively oligoribonucleotides beginning from pentaribonucleotides. From the random mixtures of penta-, hexa-, hepta-, octa-, nona- and decaribonucleotides the hepta- and octaribonucleotides are bound most efficiently. The T7 RNA-polymerase bound oligoribonucleotides can be completely extracted from the random mixture by the addition of the redundant enzyme amounts. As far as E. coli RNA-polymerase and T7 RNA-polymerase do not compete for the oligoribonucleotides the conclusion is made that they bind different oligoribonucleotides. The addition of the T7 DNA to the previously formed T7 RNA-polymerase--heptaribonucleotide complex competitively displace the heptaribonucleotides from the complex; the competitive effect of T4 DNA is very low. The data suggest that the oligoribonucleotides which are selectively bound by the RNA-polymerase are attached to the enzyme site responsible for the interaction with the promotor.
先前的研究表明,与特定长度的随机寡核苷酸混合物一起孵育的大肠杆菌RNA聚合酶会结合某些长度大于或等于5个核苷酸的寡核糖核苷酸。所呈现的数据表明,T7噬菌体诱导的RNA聚合酶(T7 RNA聚合酶)也从五核糖核苷酸开始选择性地结合寡核糖核苷酸。在五、六、七、八、九和十核糖核苷酸的随机混合物中,七核糖核苷酸和八核糖核苷酸的结合效率最高。通过添加过量的酶,可以从随机混合物中完全提取出与T7 RNA聚合酶结合的寡核糖核苷酸。由于大肠杆菌RNA聚合酶和T7 RNA聚合酶不会竞争寡核糖核苷酸,因此得出结论,它们结合不同的寡核糖核苷酸。将T7 DNA添加到先前形成的T7 RNA聚合酶 - 七核糖核苷酸复合物中会竞争性地将七核糖核苷酸从复合物中置换出来;T4 DNA的竞争作用非常低。数据表明,被RNA聚合酶选择性结合的寡核糖核苷酸附着在负责与启动子相互作用的酶位点上。