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[由多核苷酸磷酸化酶合成的RNA在经甲苯处理的大肠杆菌细胞中的核苷酸组成]

[Nucleotide composition of RNA, synthesized by polynucleotide phosphorylase, in toluene-treated cells of Escherichia coli].

作者信息

Rebrov L B

出版信息

Vopr Med Khim. 1975 Jul-Aug;21(4):409-17.

PMID:766393
Abstract

On incubation of cells of E. coli B and MRE 600 (logariphmic phase of growth), treated with toluene in presence of a mixture 14C-nucleoside-5'-diphosphates, Mg2+ or Mn2+ and tris HCl buffer pH 8.0, intracellular synthesis of heteropolyribonucleotide was observed. The synthesis was catalyzed by polynucleotide phosphorylase (PNPase, E. C. 2.7.7.8). An increase in GDP concentration in the medium distinctly decreased the incorporation of other NDP into the polymer (poly-AGUC). If the ratio of ADP, UDP, CDP, GDP in the medium was 1:1:1:0.2, the composition of nitrogenous bases in the heteropolymer produced reflected completely the NDP concentrations in the incubation mixture. Addition of different amino acids (1-lysine, 1-histidine, glycine, 1-phenylalanine) and their mixtures stimulated poly-AGUC synthesis markedly and caused an appreciable alteration in the nucleotide composition of the poly-AGUC synthesized. This phenomenon resembled the effect of amino acids on the activity of partially purified PNPase and on RNA synthesis, catalized by the enzyme in vitro. These data suggest that in bacterial cell, i. e. in vivo, PNPase synthesizes specific RNA polyribonucleotide sequences, participating in protein synthesis or in its regulation.

摘要

将处于对数生长期的大肠杆菌B和MRE 600细胞,在含有14C - 核苷 - 5'-二磷酸、Mg2+或Mn2+以及pH 8.0的三羟甲基氨基甲烷盐酸盐缓冲液的混合物存在下,用甲苯处理后进行孵育,观察到细胞内有杂多聚核糖核苷酸的合成。该合成由多核苷酸磷酸化酶(PNPase,E.C. 2.7.7.8)催化。培养基中GDP浓度的增加明显降低了其他NDP掺入聚合物(聚AGUC)的量。如果培养基中ADP、UDP、CDP、GDP的比例为1:1:1:0.2,所产生的杂聚物中含氮碱基的组成完全反映了孵育混合物中NDP的浓度。添加不同的氨基酸(1 - 赖氨酸、1 - 组氨酸、甘氨酸、1 - 苯丙氨酸)及其混合物显著刺激了聚AGUC的合成,并导致所合成的聚AGUC的核苷酸组成发生明显改变。这种现象类似于氨基酸对部分纯化的PNPase活性以及该酶在体外催化的RNA合成的影响。这些数据表明,在细菌细胞中,即在体内,PNPase合成参与蛋白质合成或其调控的特定RNA多聚核糖核苷酸序列。

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