Krzyzek R A, Rogers P
J Bacteriol. 1976 Apr;126(1):365-76. doi: 10.1128/jb.126.1.365-376.1976.
The chemical stability of argECBH messenger ribonucleic acid (mRNA) produced by Escherichia coli was found to be unaltered during steady-state repression by arginine. During extreme arginine deprivation, the increase in argECBH mRNA stability was related to general effects of amino acid starvation on mRNA stability. Thus a mechanism whereby argECBH gene expression is regulated by altering the decay rate of this mRNA is not consistent with our data. Sucrose gradient analysis followed by hybridization revealed that both heavy (14S) and light (8S) components of argECBH mRNA were produced by cells of E. coli K-12 grown without arginine, whereas predominantly light (8S) mRNA was produced by cells grown with arginine. A functional argR gene and the EC portion of the argECBH cluster were found essential for the arginine restriction of heavy-mRNA production. Experiments suggest that light argECBH mRNA did not arise from heavy message, and 8u% of both light and heavy mRNA was found bound to ribosomes. The data appear most consistent with the notion that a second site of control by arginine regulates the amounts of light and heavy arginine mRNA in the cell either by early termination of transcription or by endonucleolytic processing. Consideration of these data in conjunction with those of the accompanying report (Krzyzek and Rogers, 1976) permits the tentative conclusion that light argECBH mRNA is not translated into active enzymes and is thus responsible for the discrepancy between the high content of hybridizable mRNA and low rates of enzyme synthesis found during arginine repression.
大肠杆菌产生的精氨酸酶基因簇(argECBH)信使核糖核酸(mRNA)的化学稳定性在精氨酸稳态抑制过程中未发生改变。在精氨酸极度缺乏时,argECBH mRNA稳定性的增加与氨基酸饥饿对mRNA稳定性的总体影响有关。因此,通过改变该mRNA的降解速率来调节argECBH基因表达的机制与我们的数据不一致。蔗糖梯度分析后进行杂交显示,在无精氨酸条件下生长的大肠杆菌K-12细胞产生了argECBH mRNA的重(14S)、轻(8S)两种组分,而在有精氨酸条件下生长的细胞主要产生轻(8S)mRNA。发现功能性的argR基因和argECBH基因簇的EC部分对于重mRNA产生的精氨酸限制至关重要。实验表明,轻的argECBH mRNA并非由重的mRNA产生,并且发现80%的轻、重mRNA都与核糖体结合。这些数据似乎最符合这样一种观点,即精氨酸的第二个控制位点通过转录的早期终止或核酸内切加工来调节细胞中轻、重精氨酸mRNA的量。将这些数据与随附报告(Krzyzek和Rogers,1976)的数据相结合,可以初步得出结论,轻的argECBH mRNA不会被翻译成活性酶,因此导致了在精氨酸抑制过程中可杂交mRNA含量高与酶合成速率低之间的差异。