McMillen J, Nazario M, Jensen T
J Bacteriol. 1974 Jan;117(1):242-51. doi: 10.1128/jb.117.1.242-251.1974.
Transfer ribonucleic acid (tRNA) from exponentially growing cells (trophozoites) and from precysts of Acanthamoeba castellanii were examined by reversed-phase column (RPC-2) chromatography. This system gave excellent resolution of isoaccepting species of tRNA. The tRNAs for 12 amino acids were studied. A comparison of trophozoite and precyst tRNA elution profiles revealed no apparent differences in the number of isoaccepting species of alanyl-, arginyl-, asparaginyl-, glycyl-, leucyl-, lysyl-, methionyl-, phenylalanyl-, tryptophanyl-, or valyl-tRNAs. Seryl-tRNAs from trophozoites were eluted as three components, whereas precyst seryl-tRNAs were eluted as only two components. Precharged trophozoite and precyst isoleucyl-tRNAs were both eluted as single components; however, post-chromatography charging of trophozoite tRNA resulted in three components of activity for tRNA(Ile) and only one component for precyst tRNA(Ile). None of the observed changes could be attributed to differences in synthetases or to the presence of altered tRNA lacking the CCA terminus or partially degraded by nucleases. The possible significance of these observations is discussed.
采用反相柱(RPC - 2)色谱法对指数生长期细胞(滋养体)和卡氏棘阿米巴包囊前期的转运核糖核酸(tRNA)进行了检测。该系统对tRNA的同功受体种类具有出色的分离效果。研究了12种氨基酸的tRNA。滋养体和包囊前期tRNA洗脱图谱的比较显示,丙氨酰 -、精氨酰 -、天冬酰胺酰 -、甘氨酰 -、亮氨酰 -、赖氨酰 -、甲硫氨酰 -、苯丙氨酰 -、色氨酰 - 或缬氨酰 - tRNA的同功受体种类数量没有明显差异。滋养体的丝氨酰 - tRNA洗脱为三个组分,而包囊前期的丝氨酰 - tRNA仅洗脱为两个组分。预负载的滋养体和包囊前期异亮氨酰 - tRNA均洗脱为单一组分;然而,滋养体tRNA色谱后负载导致tRNA(Ile)的活性有三个组分,而包囊前期tRNA(Ile)只有一个组分。观察到的变化均不能归因于合成酶的差异,也不能归因于缺乏CCA末端或被核酸酶部分降解的tRNA变体的存在。讨论了这些观察结果可能具有的意义。