Iwanicka-Nowicka R, Hryniewicz M M
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
Gene. 1995 Dec 1;166(1):11-7. doi: 10.1016/0378-1119(95)00606-8.
The cbl (cysB-like) gene has been identified in Escherichia coli. The analysis of the cloned cbl sequence revealed strict homology to an ORF of unknown function found initially in Klebsiella aerogenes [Schwacha and Bender, J. Bacteriol. 175 (1993) 2107-2115]. The predicted Cbl protein has structural features of the LysR family of transcriptional activators. It is also strongly similar to the CysB protein, the activator of the cys regulon. The position of cbl on the Ec physical map has been established at a 2070-kb (43.5 min) region between asnU and asnV. The gene is expressed in vivo as a 1-kb monocistronic transcript starting from one major transcription start point. Unexpectedly, the in vivo expression of cbl has shown dependence on CysB, belonging to the same family of proteins. The promoter region of cbl binds purified CysB protein in a manner similar to other CysB-responsive promoters. A cbl disruption mutant was constructed by insertion of a KmR gene cartridge into the ORF on the chromosome. Phenotypes related to cbl expression suggest the involvement of the gene in an accessory regulatory circuit within the cys regulon engaging, in the last step, the function of the cysM gene encoding O-acetylserine (thiol)-lyase B.
cbl(类cysB)基因已在大肠杆菌中被鉴定出来。对克隆的cbl序列分析表明,它与最初在产气克雷伯菌中发现的一个功能未知的开放阅读框(ORF)具有严格的同源性[施瓦查和本德,《细菌学杂志》175(1993)2107 - 2115]。预测的Cbl蛋白具有转录激活因子LysR家族的结构特征。它也与CysB蛋白(半胱氨酸调节子的激活因子)非常相似。已确定cbl在大肠杆菌物理图谱上的位置在asnU和asnV之间的一个2070 kb(43.5分钟)区域。该基因在体内作为一个1 kb的单顺反子转录本表达,起始于一个主要转录起始点。出乎意料的是,cbl在体内的表达显示出对CysB的依赖性,CysB属于同一蛋白家族。cbl的启动子区域以类似于其他CysB应答启动子的方式结合纯化的CysB蛋白。通过将一个KmR基因盒插入染色体上的开放阅读框构建了一个cbl缺失突变体。与cbl表达相关的表型表明该基因参与了半胱氨酸调节子内的一个辅助调节回路,该回路在最后一步涉及编码O - 乙酰丝氨酸(硫醇)裂合酶B的cysM基因的功能。