Babkina G T, Karpova G G, Berzin' V A, Gren E Ia, Tsielens I E
Bioorg Khim. 1983 Nov;9(11):1535-43.
Hexaribouridylic acid, prepared by digestion of poly(U) with cobra venom endonuclease, and trinucleotide AUG synthesized chemically by triester approach were joined by RNA-ligase to yield a nonaribonucleotide AUGU6 bearing the initiation codon at its 5'-terminus. 2',3'-O-(4-[N-(2-chloro(or hydroxy) ethyl-N-Methylamino])- benzylidene residues were introduced at the 3'-terminus of oligonucleotide AUGU6 and its benzylidene derivatives AUGU6CHRCl or AUGU6CHROH were obtained. The mRNA analogs synthesized were tested for their template activity in the formation of 70S initiation complex. AUGU6, AUGU6CHRC1 and AUGU6CHROH were shown to stimulate factor-dependent binding of fMet-tRNA to ribosomes. The effect of benzylidene fragment on the template activity of AUGU6CHROH in the course of translation process was studied. It was shown that AUGU6CHROH stimulates synthesis of di- and tripeptides with the same efficiency as AUGU6.
用眼镜蛇毒核酸内切酶消化聚(U)制备的六聚核糖核苷酸,与通过三酯法化学合成的三核苷酸AUG,经RNA连接酶连接,得到在其5'-末端带有起始密码子的九聚核糖核苷酸AUGU6。在寡核苷酸AUGU6的3'-末端引入2',3'-O-(4-[N-(2-氯(或羟基)乙基-N-甲基氨基])-亚苄基残基,得到其亚苄基衍生物AUGU6CHRCl或AUGU6CHROH。对合成的mRNA类似物在形成70S起始复合物中的模板活性进行了测试。结果表明,AUGU6、AUGU6CHRC1和AUGU6CHROH能刺激fMet-tRNA与核糖体的因子依赖性结合。研究了亚苄基片段在翻译过程中对AUGU6CHROH模板活性的影响。结果表明,AUGU6CHROH刺激二肽和三肽合成的效率与AUGU6相同。