Zacharias M, Wagner R
Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin, FRG.
Nucleic Acids Res. 1987 Oct 26;15(20):8235-48. doi: 10.1093/nar/15.20.8235.
A number of deletions have been constructed within the leader region of the rrnB operon from E. coli. The deletions remove a potential transcription terminator structure downstream from an antitermination recognition sequence (Box A), which precedes the structural gene for the 16S RNA. Cells harbouring plasmids, where the terminator structure was deleted, partially or totally, showed a reduction in growth rate under minimal growth conditions. Measurement of the ribosomal RNA synthesis rates of such cells determined by pulselabeling and hybridisation to appropriate DNA probes, showed that the amount of the more distally located 23S RNA was reduced compared to the promoter-proximal 16S RNA. This polarity in transcription, resulting in a non-stoichiometric synthesis of the ribosomal RNAs, is most likely the result of a defective antitermination. The reduction in the amount of 23S RNA in such cells is compensated for by an increase in the overall ribosomal RNA synthesis, in concordance with the ribosomal RNA feedback regulation model. The accumulation of transcripts of the tRNAGlu2 gene, coded in the spacer region between the 16S and 23S RNA genes, in cells with an altered rRNA stoichiometry supports this interpretation.
已经在来自大肠杆菌的rrnB操纵子的前导区域内构建了一些缺失。这些缺失去除了位于抗终止识别序列(A框)下游的一个潜在转录终止子结构,该序列位于16S RNA的结构基因之前。携带质粒的细胞,其中终止子结构被部分或完全删除,在基本生长条件下显示出生长速率降低。通过脉冲标记和与合适的DNA探针杂交来测定此类细胞的核糖体RNA合成速率,结果表明,与启动子近端的16S RNA相比,位置更远的23S RNA的量减少了。这种转录极性导致核糖体RNA的非化学计量合成,很可能是抗终止缺陷的结果。此类细胞中23S RNA量的减少通过核糖体RNA总体合成的增加得到补偿,这与核糖体RNA反馈调节模型一致。在rRNA化学计量改变的细胞中,编码在16S和23S RNA基因之间间隔区的tRNAGlu2基因转录本的积累支持了这一解释。