Harms E, Umbarger H E
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.
J Bacteriol. 1987 Dec;169(12):5668-77. doi: 10.1128/jb.169.12.5668-5677.1987.
Leucine participates in multivalent repression of the Serratia marcescens ilvGMEDA operon by attenuation (J.-H. Hsu, E. Harms, and H.E. Umbarger, J. Bacteriol. 164:217-222, 1985), although there is only one single leucine codon that could be involved in this type of control. This leucine codon is the rarely used CUA. The contribution of this leucine codon to the control of transcription by attenuation was examined by replacing it with the commonly used leucine codon CUG and with a nonregulatory proline codon, CCG. These changes left intact the proposed secondary structure of the leader. The effects of the codon changes were assessed by placing the mutant leader regions upstream of the ilvGME structural genes or the cat gene and measuring acetohydroxy acid synthase II, transaminase B, or chloramphenicol acetyltransferase activities in cells grown under limiting and repressing conditions. The presence of the common leucine codon in place of the rare leucine codon reduced derepression by about 70%. Eliminating the leucine codon by converting it to proline abolished leucine control. Furthermore, a possible context effect of the adjacent upstream serine codon on leucine control was examined by changing it into a glycine codon.
亮氨酸通过衰减作用参与对粘质沙雷氏菌ilvGMEDA操纵子的多价阻遏(J.-H. 许、E. 哈姆斯和H.E. 昂巴格,《细菌学杂志》164:217 - 222,1985年),尽管在这种调控类型中可能涉及的亮氨酸密码子只有一个。这个亮氨酸密码子是很少使用的CUA。通过将其替换为常用的亮氨酸密码子CUG和一个非调控性的脯氨酸密码子CCG,研究了这个亮氨酸密码子对衰减转录调控的贡献。这些变化使前导序列的二级结构保持完整。通过将突变的前导序列区域置于ilvGME结构基因或cat基因的上游,并在限制和阻遏条件下生长的细胞中测量乙酰羟酸合酶II、转氨酶B或氯霉素乙酰转移酶的活性,评估密码子变化的影响。用常见的亮氨酸密码子取代罕见的亮氨酸密码子后,去阻遏作用降低了约70%。将亮氨酸密码子转换为脯氨酸从而消除该密码子后,亮氨酸调控作用消失。此外,通过将相邻的上游丝氨酸密码子改变为甘氨酸密码子,研究了其对亮氨酸调控可能的上下文效应。