Harms E, Higgins E, Chen J W, Umbarger H E
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.
J Bacteriol. 1988 Oct;170(10):4798-807. doi: 10.1128/jb.170.10.4798-4807.1988.
The hypothesis that translation of the ilvD and ilvA genes of Escherichia coli may be linked has been examined in strains in which lacZ-ilvD protein fusions are translated in all three reading frames with respect to ilvD. In these strains, the nucleotide sequence was altered to obtain premature termination of ilvD translation, and in one strain translation termination of ilvD DNA occurred two bases downstream of the ilvA initiation codon. In the wild-type strain, the ilvD translation termination site was located two bases upstream of the ilvA start codon. In each of the mutant strains, expression of ilvA, as determined by the level of threonine deaminase activity, was strikingly lower than in the wild-type strain. The data suggest that expression of ilvD and ilvA is translationally coupled. By inserting a promoterless cat gene downstream of ilvA, it was shown that the differences in enzyme activity were not the result of differences in the amount of ilvA mRNA produced.
大肠杆菌ilvD和ilvA基因的翻译可能存在关联这一假说,已在一些菌株中进行了研究,在这些菌株中,lacZ-ilvD蛋白融合体相对于ilvD在所有三个阅读框中进行翻译。在这些菌株中,改变核苷酸序列以实现ilvD翻译的提前终止,并且在一个菌株中,ilvD DNA的翻译终止发生在ilvA起始密码子下游两个碱基处。在野生型菌株中,ilvD翻译终止位点位于ilvA起始密码子上游两个碱基处。在每个突变菌株中,通过苏氨酸脱氨酶活性水平测定的ilvA表达,显著低于野生型菌株。数据表明ilvD和ilvA的表达在翻译上是偶联的。通过在ilvA下游插入一个无启动子的cat基因,结果表明酶活性的差异不是由产生的ilvA mRNA量的差异导致的。