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在食菌蛭弧菌109J周质内生长期间大肠杆菌脱氧核糖核酸的调控降解

Regulated breakdown of Escherichia coli deoxyribonucleic acid during intraperiplasmic growth of Bdellovibrio bacteriovorus 109J.

作者信息

Rosson R A, Rittenberg S C

出版信息

J Bacteriol. 1979 Nov;140(2):620-33. doi: 10.1128/jb.140.2.620-633.1979.

Abstract

During growth of Bdellovibrio bacteriovorus on [2-14C]deoxythymidine-labeled Escherichia coli, approximately 30% of the radioactivity was released to the culture fluid as nucleoside monophosphates and free bases; the remainder was incorporated by the bdellovibrio. By 60 min after bdellovibrio attack, when only 10% of the E. coli deoxyribonucleic acid (DNA) had been solubilized, the substrate cell DNA was degraded to 5 X 10(5)-dalton fragments retained within the bdelloplast. Kinetic studies showed these fragments were formed as the result of sequential accumulation of single- and then double-strand cuts. DNA fragments between 2 X 10(3) and 5 X 10(5) daltons were never observed. Chloramphenicol, added at various times after initiation of bdellovibrio intraperiplasmic growth on normal or on heated E. coli, which have inactivated deoxyribonucleases, inhibited further breakdown and solubilization of substrate cell DNA. Analysis of these intraperiplasmic culture deoxyribonuclease activities showed that bdellovibrio deoxyribonucleases are synthesized while E. coli nucleases are inactivated. It is concluded that continuous and sequential synthesis of bdellovibrio deoxyribonucleases of apparently differing specificities is necessary for complete breakdown and solubilization of substrate cell DNA, and that substrate cell deoxyribonucleases are not involved in any significant way in the degradation process.

摘要

在噬菌蛭弧菌以[2-¹⁴C]脱氧胸苷标记的大肠杆菌为食生长期间,约30%的放射性以核苷单磷酸和游离碱基的形式释放到培养液中;其余部分则被蛭弧菌摄取。在蛭弧菌攻击后60分钟,当只有10%的大肠杆菌脱氧核糖核酸(DNA)被溶解时,底物细胞DNA已被降解为保留在蛭弧体中的5×10⁵道尔顿片段。动力学研究表明,这些片段是单链切割然后双链切割顺序积累的结果。从未观察到2×10³至5×10⁵道尔顿之间的DNA片段。在噬菌蛭弧菌在正常或加热的大肠杆菌上进行周质内生长开始后的不同时间添加氯霉素,这些大肠杆菌的脱氧核糖核酸酶已失活,氯霉素抑制了底物细胞DNA的进一步分解和溶解。对这些周质内培养液的脱氧核糖核酸酶活性分析表明,噬菌蛭弧菌的脱氧核糖核酸酶是在大肠杆菌核酸酶失活时合成的。得出的结论是,连续且顺序合成明显具有不同特异性的噬菌蛭弧菌脱氧核糖核酸酶对于底物细胞DNA的完全分解和溶解是必要的,并且底物细胞脱氧核糖核酸酶在降解过程中没有任何显著作用。

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