Gavini N, Pulakat L
Russell Grimwade School of Biochemistry, University of Melbourne, Parkville, Victoria, Australia.
J Bacteriol. 1991 Aug;173(15):4904-7. doi: 10.1128/jb.173.15.4904-4907.1991.
In Escherichia coli, the expression of the pheA gene is regulated by attenuation of transcription. To study the molecular details of pheA attenuation, we introduced mutations in the pheA leader peptide coding region and analyzed their effects by using pheA promoter-lacZ gene transcription fusions (pheAp-lacZ). Mutations in the ribosome-binding site (pheAe1213) or in the translation initiation codon (pheAe24) of the pheA leader peptide coding region resulted in superattenuation of pheA expression. However, the presence of a stop codon upstream to the tandem phenylalanine codons (pheAe3334) led to an increase in the basal-level expression of pheA. This increase was further enhanced in the presence of prfA release factor mutant. The level of pheA expression in all three mutants was the same when cells were starved for phenylalanine. These results demonstrate that efficient translation of the pheA leader peptide coding region and the position of the ribosome on the leader transcript play decisive roles in the attenuation regulation of pheA.
在大肠杆菌中,pheA基因的表达受转录衰减调控。为了研究pheA衰减的分子细节,我们在pheA前导肽编码区引入突变,并通过使用pheA启动子 - lacZ基因转录融合体(pheAp - lacZ)分析其影响。pheA前导肽编码区核糖体结合位点(pheAe1213)或翻译起始密码子(pheAe24)的突变导致pheA表达的超衰减。然而,串联苯丙氨酸密码子上游存在终止密码子(pheAe3334)导致pheA基础水平表达增加。在prfA释放因子突变体存在的情况下,这种增加进一步增强。当细胞缺乏苯丙氨酸时,所有三个突变体中pheA的表达水平相同。这些结果表明,pheA前导肽编码区的有效翻译以及核糖体在引导转录本上的位置在pheA的衰减调控中起决定性作用。