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沼泽红假单胞菌7号菌株核糖体RNA操纵子的结构、表达及产物

Structure, expression and products of the ribosomal RNA operons of Rhodopseudomonas palustris No. 7.

作者信息

Zahn K, Inui M, Yukawa H

机构信息

Research Institute of Innovative Technology for the Earth, Soraku, Kyoto, Japan.

出版信息

Mol Genet Genomics. 2001 Jul;265(5):778-90. doi: 10.1007/s004380100466.

Abstract

Rhodopseudomonas palustris strains carry one or two ribosomal rRNA operons, and those with duplicated rrn operons grow faster. The two rrn operons in R. palustris No. 7 are virtually identical over a 54,70-bp stretch containing the genes for 16S rRNA, tRNAile, tRNAala, 23S rRNA and 5S rRNA, as well as the intergenic spacers and part of the extragenic spacer. In R. palustris, unlike most bacteria with multiple rrn operons, the putative promoter sequences of the two operons are highly diverged, suggesting possible functional differentiation. By simultaneous primer-extension analysis of both pre-rRNAs, we detected a two-fold higher level of expression from rrnA under photoautotrophic conditions. Alteration of the conditions of growth leads to changes in the relative levels of expression of the two operons. Within the 5,470-bp segment, only two sequence differences are found between the 23S rRNA genes; one is at the center of the 23S rRNA molecule and affects a site of unknown function, and the other is within or immediately adjacent to sequences involved in processing of the 5' 23S rRNA IVS. In vitro processing of 5' IVS-containing 23S rRNA precursors from each operon does not reveal any detectable difference between them. The 5' ends of the mature 16S, 23S, and 5S rRNAs were determined by primer-extension analysis, and the 3' end of 23S rRNA was determined by RNA linker ligation-mediated cDNA cloning. The 5' and 3' ends of the R. palustris 23S rRNA molecule are extensively processed, suggesting that, unlike the situation in the established eubacterial model, these ends cannot basepair.

摘要

沼泽红假单胞菌菌株携带一个或两个核糖体rRNA操纵子,具有重复rrn操纵子的菌株生长更快。沼泽红假单胞菌7号菌株中的两个rrn操纵子在一段5470 bp的片段上几乎完全相同,该片段包含16S rRNA、tRNAile、tRNAala、23S rRNA和5S rRNA的基因,以及基因间隔区和部分基因外间隔区。在沼泽红假单胞菌中,与大多数具有多个rrn操纵子的细菌不同,这两个操纵子的推定启动子序列高度不同,表明可能存在功能分化。通过对两种前体rRNA同时进行引物延伸分析,我们发现在光合自养条件下rrnA的表达水平高两倍。生长条件的改变会导致两个操纵子相对表达水平的变化。在5470 bp的片段内,23S rRNA基因之间仅发现两个序列差异;一个位于23S rRNA分子的中心,影响一个功能未知的位点,另一个位于参与5' 23S rRNA IVS加工的序列内或紧邻该序列。对每个操纵子含5' IVS的23S rRNA前体进行体外加工,未发现它们之间有任何可检测到的差异。通过引物延伸分析确定了成熟16S、23S和5S rRNA的5'末端,并通过RNA接头连接介导的cDNA克隆确定了23S rRNA的3'末端。沼泽红假单胞菌23S rRNA分子的5'和3'末端经过广泛加工,这表明与已建立的真细菌模型不同,这些末端不能形成碱基对。

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