Juarez H, Skjold A C, Hedgcoth C
J Bacteriol. 1975 Jan;121(1):44-54. doi: 10.1128/jb.121.1.44-54.1975.
When treated with chloramphenicol, Escherichia coli 15T minus produces two new species (IV and V) of transfer ribonucleic acid specific for phenylalanine in addition to the major normal species (II) and two minor normal species (I and III), which are seen as distinct components upon fractionation by chromatography on columns of benzoylated diethylaminoethyl-cellulose. Species IV is produced when cells are grown in iron-deficient medium and is, therefore, probably deficient in the 2-methylthio modification of N-6-(delta-2-isopentenyl) adenosine. A new minor species (Va) also appears under those conditions. All of the new components elute earlier than the major normal species. Addition of chloramphenicol to iron-deficient cells leads to the production of species V, and that production is blocked by rifampin, as is the production of species IV. Thus, species IV and V appear to be transcriptional products. Although E. coli 15T minus appears to be rel plus, starvation for methionine or cysteine leads to the accumulation of species IV (without addition of chloramphenicol); rifampin blocks the accumulation. Species V is still produced on addition of chloramphenicol to starved cultures. Starvation for arginine or tryptophan does not alter the chromatographic profile from the normal case. Treatment with permanganate indicates that species II and IV contain isopentenyladenosine but that species V does not. Species V appears to be deficient in both isopentenyl and methylthio modifications of adenosine and perhaps at least one other modification, because removing the isopentenyl moiety from adenosine does not convert species IV into species V, but converts it into species Va. A precursor relationship among species V, VI, and II is suggested by following the chromatographic profile of phenylalanine transfer ribonucleic acid during recovery of E. coli from treatment with chloramphenicol; the various species increase and decrease in a sequential manner.
用氯霉素处理时,大肠杆菌15T减株除产生主要的正常种类(II)和两个次要的正常种类(I和III)外,还产生两种新的苯丙氨酸特异性转移核糖核酸种类(IV和V),这些种类在苯甲酰化二乙氨基乙基纤维素柱上进行色谱分离时表现为不同的组分。种类IV是细胞在缺铁培养基中生长时产生的,因此可能缺乏N-6-(δ-2-异戊烯基)腺苷的2-甲硫基修饰。在这些条件下还会出现一种新的次要种类(Va)。所有新组分的洗脱都早于主要的正常种类。向缺铁细胞中添加氯霉素会导致种类V的产生,而利福平会阻断其产生,种类IV的产生也会被利福平阻断。因此,种类IV和V似乎是转录产物。尽管大肠杆菌15T减株似乎是rel加型,但蛋氨酸或半胱氨酸饥饿会导致种类IV的积累(无需添加氯霉素);利福平会阻断这种积累。向饥饿培养物中添加氯霉素时仍会产生种类V。精氨酸或色氨酸饥饿不会改变色谱图谱与正常情况的差异。用高锰酸盐处理表明,种类II和IV含有异戊烯基腺苷,而种类V不含。种类V似乎在腺苷的异戊烯基和甲硫基修饰以及可能至少一种其他修饰方面存在缺陷,因为从腺苷中去除异戊烯基部分不会将种类IV转化为种类V,而是将其转化为种类Va。在大肠杆菌从氯霉素处理中恢复过程中跟踪苯丙氨酸转移核糖核酸的色谱图谱,提示了种类V、VI和II之间的前体关系;各种类以连续的方式增加和减少。