Blum P H
Department of Biochemistry, University of California, Berkeley 94720.
J Bacteriol. 1988 Nov;170(11):5125-33. doi: 10.1128/jb.170.11.5125-5133.1988.
Salmonella typhimurium miaA mutants lacking the tRNA base modification cis-2-methylthioribosylzeatin (ms2io6A) were examined and found to be sensitive to a variety of chemical oxidants and unable to grow aerobically at 42 degrees C in a defined medium. Leucine supplementation suppressed both of these phenotypes, suggesting that leucine synthesis was defective. Intracellular levels of leucine decreased 40-fold in mutant strains after a shift from 30 to 42 degrees C during growth, and expression of a leu-lacZ transcriptional fusion ceased. Steady-state levels of leu mRNA were also significantly reduced during growth at elevated temperatures. Failure of miaA mutant leu-lacZ expression to be fully derepressed during L-leucine limitation at 30 degrees C and suppression of the miaA mutation by a mutation in the S. typhimurium leu attenuator suggests that translational control of the transcription termination mechanism regulating leu expression is defective. Since the S. typhimurium miaA mutation was also suppressed by the Escherichia coli leu operon in trans, phenotypic differences between E. coli and S. typhimurium miaA mutants may result from a difference between their respective leu operons.
对缺乏tRNA碱基修饰顺式-2-甲基硫代核糖基玉米素(ms2io6A)的鼠伤寒沙门氏菌miaA突变体进行了检测,发现它们对多种化学氧化剂敏感,并且在限定培养基中于42℃无法进行有氧生长。补充亮氨酸可抑制这两种表型,表明亮氨酸合成存在缺陷。在生长过程中从30℃转移至42℃后,突变菌株中亮氨酸的细胞内水平下降了40倍,并且leu-lacZ转录融合体的表达停止。在高温生长期间,leu mRNA的稳态水平也显著降低。在30℃的L-亮氨酸限制期间,miaA突变体leu-lacZ表达未能完全去阻遏,以及鼠伤寒沙门氏菌亮氨酸弱化子中的突变对miaA突变的抑制作用,表明调节亮氨酸表达的转录终止机制的翻译控制存在缺陷。由于大肠杆菌的leu操纵子也能反式抑制鼠伤寒沙门氏菌的miaA突变,大肠杆菌和鼠伤寒沙门氏菌miaA突变体之间的表型差异可能源于它们各自leu操纵子之间的差异。