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大肠杆菌突变体中核糖体RNA大前体的合成。

Synthesis of a large precursor to ribosomal RNA in a mutant of Escherichia coli.

作者信息

Nikolaev N, Silengo L, Schlessinger D

出版信息

Proc Natl Acad Sci U S A. 1973 Dec;70(12):3361-5. doi: 10.1073/pnas.70.12.3361.

Abstract

A mutant of E. coli, isolated by Kindler and Hofschneider as a strain defective in RNase III activity, forms a 30S precursor of ribosomal RNA ("30S pre-rRNA"). The half-life of the 30S pre-rRNA in growing cells at 30 degrees , estimated by the rate of specific (3)[H]uridine incorporation, is about 1 min. In rifampicin-treated cells, the RNA is metabolized to mature rRNA with a half-life of about 2 min. The 30S pre-rRNA has been highly purified. DNA-RNA hybridization tests demonstrate that it contains both 16S and 23S rRNA sequences. Also, in cultures treated with rifampicin, the cleavage products of radioactive 30S pre-rRNA include 25S and 17.5S RNA species, destined to becomes 23S and 16S rRNA. Thus, each 30S chain probably contains one 16S and one 23S RNA sequence, as well as additional sequences. Two independent techniques indicate that the additional portions account for about 27% of the total lenght: (1) By comparison to the sedimentation rate and electrophoretic mobility of marker RNAs, the 30S pre-RNA has an apparent molecular weight of 2.3 x 10(6) +/- 5%, or 28% more than the sum of 16S and 23S rRNA; (2) 27% of the 30S pre-rRNA is not competed away from hybridization by mature 16S and 23S rRNA.Thus, bacteria appear to make a pre-rRNA similar in some respects to that observed in eukaryotes; though in normal E. coli cells, the pre-rRNA is ordinarily cleaved endonucleolytically during its formation.

摘要

金德勒和霍夫施奈德分离出的一株大肠杆菌突变体,其核糖核酸酶III活性有缺陷,能形成核糖体RNA的30S前体(“30S前体rRNA”)。通过特定的(3)[H]尿苷掺入速率估算,30S前体rRNA在30摄氏度的生长细胞中的半衰期约为1分钟。在利福平处理的细胞中,该RNA代谢为成熟rRNA,半衰期约为2分钟。30S前体rRNA已被高度纯化。DNA-RNA杂交试验表明它含有16S和23S rRNA序列。此外,在用利福平处理的培养物中,放射性30S前体rRNA的切割产物包括25S和17.5S RNA种类,它们将成为23S和16S rRNA。因此,每个30S链可能包含一个16S和一个23S RNA序列,以及其他序列。两种独立技术表明,这些额外部分约占总长度的27%:(1)与标记RNA的沉降速率和电泳迁移率相比,30S前体RNA的表观分子量为2.3×10(6)±5%,比16S和23S rRNA的总和多28%;(2)30S前体rRNA的27%不会被成熟的16S和23S rRNA从杂交中竞争掉。因此,细菌似乎产生了一种在某些方面与真核生物中观察到的前体rRNA相似的前体rRNA;尽管在正常大肠杆菌细胞中,前体rRNA通常在其形成过程中被内切核酸酶切割。

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