Bradford D S, Hacker B, Clark I
Biochem J. 1972 Mar;126(5):1057-66. doi: 10.1042/bj1261057a.
Methods were devised for the assay of tRNA methylases of rat bone. The activities of bone tRNA methylases are similar to those from other mammalian tissues. However, unlike reports on liver methylases, no inhibitors were found in the supernatant fraction from pH5 precipitate of bone extracts. The effects of vitamins A and D on the methylation of tRNA by cell-free extracts of rat bone were studied. Deficiency of either vitamin resulted in a decrease in the rate and extent of tRNA methylation, whereas the administration of vitamin A to hypovitaminotic-A rats and vitamin D to hypovitaminotic-D rats increased the rate and extent of tRNA methylation. These effects appear to be apart from changes in ribonuclease activity or in concentrations of calcium or magnesium. No evidence of inhibitors of tRNA methylases was found in bone extracts from vitamin-deficient rats nor of activators in bone extracts from deficient rats given vitamin A or D. The pattern of tRNA methylation under conditions of vitamin A or D deficiency was not changed, suggesting a generalized cellular deficiency. It was of significance to find that the specificity for methylation of specific bases in tRNA was different after the administration of vitamin A as contrasted with the effects of vitamin D. The possible significance of tRNA methylation to the biochemical action of the vitamins on bone is discussed.
已设计出测定大鼠骨骼中tRNA甲基化酶的方法。骨骼tRNA甲基化酶的活性与其他哺乳动物组织中的相似。然而,与关于肝脏甲基化酶的报道不同,在骨骼提取物pH5沉淀的上清液部分未发现抑制剂。研究了维生素A和D对大鼠骨骼无细胞提取物tRNA甲基化的影响。缺乏任何一种维生素都会导致tRNA甲基化的速率和程度降低,而给维生素A缺乏的大鼠补充维生素A以及给维生素D缺乏的大鼠补充维生素D,则会增加tRNA甲基化的速率和程度。这些影响似乎与核糖核酸酶活性的变化或钙或镁浓度的变化无关。在维生素缺乏大鼠的骨骼提取物中未发现tRNA甲基化酶抑制剂的证据,在给予维生素A或D的缺乏大鼠的骨骼提取物中也未发现激活剂的证据。维生素A或D缺乏条件下tRNA甲基化的模式没有改变,表明存在普遍的细胞缺陷。值得注意的是,与维生素D的作用相比,给予维生素A后tRNA特定碱基甲基化的特异性有所不同。文中讨论了tRNA甲基化对维生素在骨骼上生化作用的可能意义。