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噬菌体T4感染大肠杆菌期间宿主蛋白质合成的抑制作用。I. 宿主核糖核酸的持续合成。

Inhibition of host protein synthesis during infection of Escherichia coli by bacteriophage T4. I. Continued synthesis of host ribonucleic acid.

作者信息

Kennell D

出版信息

J Virol. 1968 Nov;2(11):1262-71. doi: 10.1128/JVI.2.11.1262-1271.1968.

Abstract

The ribonucleic acid (RNA) synthesized at specified intervals during infection of Escherichia coli K-12 by bacteriophage T4 was hybridized to denatured E. coli or T4 deoxyribonucleic acids (DNA). The reactions were performed under conditions that maximized the yield and at RNA/DNA inputs such that excess DNA sites were available for all RNA species. Most of the RNA synthesized at any time during the first 3 min of infection was host-specific. The fraction declined rapidly as infection progressed; host RNA represented about half that made between 3 and 4 min. It is unlikely that this represented RNA synthesized by bacteria that had escaped infection, as judged by the kinetics of adsorption and killing as well as by the rapid inhibition of beta-galactosidase induction after infection. The nature of the host RNA was also examined. Part of the RNA synthesized during infection of cells rendered sensitive to actinomycin was stable in the presence of this inhibitor. This RNA was essentially all host-specific and it sedimented as ribosomal and transfer RNA; most of the ribosomal RNA was incorporated into 30S and 50S ribosomes. Hybridization analyses suggested that unstable E. coli messenger RNA was also synthesized for several minutes after infection; the proportion of unstable to stable host RNA synthesized appeared to be similar in infected and uninfected cells. Thus, it is concluded that significant amounts of E. coli RNA are synthesized during the first minutes of T4 infection. Host messenger RNA initiated after infection may not be translated into enzymes; alternatively, it is conceivable that continued bacterial messenger RNA synthesis only reflects the completion of transcription of operons whose reading was initiated prior to infection.

摘要

在噬菌体T4感染大肠杆菌K-12的特定时间间隔内合成的核糖核酸(RNA)与变性的大肠杆菌或T4脱氧核糖核酸(DNA)进行杂交。反应在使产量最大化的条件下进行,且RNA/DNA输入量要保证所有RNA种类都有过量的DNA位点可供杂交。感染后最初3分钟内任何时间合成的大部分RNA都是宿主特异性的。随着感染的进行,这一比例迅速下降;宿主RNA在3到4分钟之间合成的量约为之前的一半。从吸附和杀灭动力学以及感染后β-半乳糖苷酶诱导的快速抑制情况判断,这不太可能代表未被感染的细菌合成的RNA。还对宿主RNA的性质进行了研究。在感染过程中对放线菌素敏感的细胞中合成的部分RNA在该抑制剂存在的情况下是稳定的。这种RNA基本上都是宿主特异性的,并且沉降为核糖体RNA和转移RNA;大部分核糖体RNA被整合到30S和50S核糖体中。杂交分析表明,感染后几分钟内还合成了不稳定的大肠杆菌信使RNA;在感染和未感染的细胞中,合成的不稳定宿主RNA与稳定宿主RNA的比例似乎相似。因此,可以得出结论,在T4感染的最初几分钟内合成了大量的大肠杆菌RNA。感染后起始的宿主信使RNA可能不会被翻译成酶;或者,可以想象,细菌信使RNA的持续合成仅仅反映了在感染前起始转录的操纵子转录的完成。

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