Deana A, Ehrlich R, Reiss C
Centre de Génétique Moléculaire, Laboratoire Structure et Dynamique du Génome, CNRS, F91198 Gif-sur-Yvette, France.
Nucleic Acids Res. 1998 Oct 15;26(20):4778-82. doi: 10.1093/nar/26.20.4778.
In order to test the effect of silent mutations on the regulation of gene expression, we monitored several steps of transcription and translation of the ompA gene in vivo , in which some or all codons between codons 6 and 14, frequently used in Escherichia coli , had been exchanged for infrequent synonymous codons. Northern blot analysis revealed an up to 4-fold reduction in the half-life of the mutated messengers and a >10-fold reduction in their steady-state amounts. Western blot analysis showed a 10-fold reduction in the amount of OmpA protein. Use of a system expressing a Rho-specific anti-terminator allowed us to detect a strong transcription polarity effect in the silent mutants. These results demonstrate that silent mutations can severely inhibit several steps of gene expression in E. coli and that code degeneracy is efficiently exploited in this species for setting signals for gene control and regulation.
为了测试沉默突变对基因表达调控的影响,我们在体内监测了ompA基因转录和翻译的几个步骤,其中大肠杆菌中常用的密码子6至14之间的一些或所有密码子已被替换为不常用的同义密码子。Northern印迹分析显示,突变信使的半衰期最多降低了4倍,其稳态量降低了10倍以上。Western印迹分析显示OmpA蛋白的量减少了10倍。使用表达Rho特异性抗终止子的系统使我们能够在沉默突变体中检测到强烈的转录极性效应。这些结果表明,沉默突变可严重抑制大肠杆菌中基因表达的几个步骤,并且该物种有效地利用密码简并性来设置基因控制和调节信号。