Kaga Naoko, Umitsuki Genryou, Clark David P, Nagai Kazuo, Wachi Masaaki
Department of Bioengineering, Tokyo Institute of Technology, Yokohama 226-8501, Japan.
Biochem Biophys Res Commun. 2002 Jul 5;295(1):92-7. doi: 10.1016/s0006-291x(02)00639-3.
Escherichia coli RNase G encoded by the rng gene is involved in degradation of adhE mRNA. Overproduction of the AdhE protein by rng mutants was found to depend on the genetic background of strains derived from DC272 (adhC81) or MC1061. We found that DC272 carried a point mutation in the Cra-binding site of the adhE promoter. The Cra protein encoded by the cra gene is known to act as a repressor of adhE. P1-phage-mediated transduction and lacZ fusion analysis with the mutant adhE promoter confirmed that this mutation is responsible for overproduction. On the other hand, Southern hybridization revealed that MC1061 had a 0.85-kb deletion of the cra gene. Overproduction of AdhE in the MC1061 background was reversed to the wild-type levels by introduction of a plasmid carrying the cra(+) gene. These results indicated that expression of the adhE gene was regulated transcriptionally by Cra and posttranscriptionally by RNase G.
由rng基因编码的大肠杆菌核糖核酸酶G参与adhE信使核糖核酸的降解。发现rng突变体中AdhE蛋白的过量产生取决于源自DC272(adhC81)或MC1061的菌株的遗传背景。我们发现DC272在adhE启动子的Cra结合位点携带一个点突变。已知由cra基因编码的Cra蛋白作为adhE的阻遏物起作用。用突变的adhE启动子进行P1噬菌体介导的转导和lacZ融合分析证实该突变是过量产生的原因。另一方面,Southern杂交显示MC1061有一个0.85 kb的cra基因缺失。通过引入携带cra(+)基因的质粒,MC1061背景中AdhE的过量产生恢复到野生型水平。这些结果表明adhE基因的表达受Cra转录调控和核糖核酸酶G的转录后调控。