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用d(pC-G)-A的烷基化衍生物对酵母tRNA Val 1进行互补寻址修饰。复合物中烷基化核苷酸的位置及烷基化过程。

Complementary addressed modification of yeast tRNA Val 1 with alkylating derivative of d(pC-G)-A. The positions of the alkylated nucleotides and the course of the alkylation in the complex.

作者信息

Grineva N I, Karpova G G, Kuznetsova L M, Venkstern T V, Bayev A A

出版信息

Nucleic Acids Res. 1977;4(5):1609-31. doi: 10.1093/nar/4.5.1609.

Abstract

Yeast tRNA Val 1 alkylation with 2', 3'-O-4-(N-2-chloroethyl-N-methylamino) benzylidene d(pC-G)-A proceeds at 20 degrees - 30 degrees C in the complementary complexes which are formed by d(pC-G)-A greater than RC1 binding to 3 sequences of tRNA Val 1 : psi-C-G58 in the T loop, C-G40 at the 3'-side of the anticodon loop and C-G18 in the D loop. The reaction in the complexes results in A53, I35, and psi 13 alkylation to form beta-/N-methyl-N-(formylphenyl 17 amino/ethyl-tRNA Val 1 with the relative rate constants of the alkylation that are 3 or 2 orders of magnitude higher than that for the alkylation without a complex formation. It is the third nucleotide from the 5'-terminus of the binding site of the modifying agent that is subjected to alkylation in the t RNA Val 1. The course of the alkylation does not depend on the possible base pairing of the 3'-terminal nucleotide of the reagent. The extent of the reagent binding and the relative rate constants of the alkalytion in the complexes indicate the following order of the complex stability: (psi-C-G58) greater than (CO-G40) approximately (C-G18) at 20 degrees and (psi-C-G58) greater than (C-G40) greater than (C-G18) at 30 degrees.

摘要

酵母tRNA Val 1与2', 3'-O-4-(N-2-氯乙基-N-甲氨基)亚苄基d(pC-G)-A的烷基化反应在20℃至30℃下于互补复合物中进行,该复合物由d(pC-G)-A大于RC1与tRNA Val 1的3个序列结合形成:T环中的ψ-C-G58、反密码子环3'-侧的C-G40以及D环中的C-G18。复合物中的反应导致A53、I35和ψ13烷基化,形成β-/N-甲基-N-(甲酰苯基17氨基/乙基)-tRNA Val 1,其烷基化的相对速率常数比未形成复合物时的烷基化速率常数高3或2个数量级。在t RNA Val 1中发生烷基化的是修饰剂结合位点5'-末端的第三个核苷酸。烷基化过程不依赖于试剂3'-末端核苷酸可能的碱基配对。复合物中试剂的结合程度和烷基化的相对速率常数表明复合物稳定性的顺序如下:20℃时(ψ-C-G58)大于(CO-G40)约等于(C-G18),30℃时(ψ-C-G58)大于(C-G40)大于(C-G18)。

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