Quivy J P, Chroboczek J
European Molecular Biology Laboratory, URA 1333, Grenoble, France.
Biochimie. 1991 Oct;73(10):1269-73. doi: 10.1016/0300-9084(91)90087-h.
The effect of aminoacylation of the tRNA-like end of brome mosaic virus RNA during in vitro protein synthesis and in vitro viral encapsidation was investigated. The components of the homologous system were: BMV RNA, wheat germ cell-free protein synthesizing system and pure tyrosyl-tRNA synthetase from wheat germ. During in vitro protein synthesis directed with tyrosylated as well as non-tyrosylated BMV RNA, no differences were observed in the amount and in the class of polypeptides formed neither in the velocity of the translation reaction. Excess active TyrRS was added during in vitro translation, without modifying the translation efficiency. BMV RNA and active TyrRS were preincubated prior to translation in order to interact without the translation system components and then subjected to translation in vitro. Similar results were obtained when BMV RNA was preincubated with inactive TyrRS or BSA. These results indicate that the aminoacylation of BMV RNA has no pronounced effect on viral protein synthesis in vitro. During BMV RNA encapsidation either tyrosylated or non-tyrosylated BMV RNA 4 could be encapsidated in a similar way.
研究了在体外蛋白质合成和体外病毒衣壳化过程中，雀麦花叶病毒RNA类tRNA末端的氨酰化作用。同源系统的组成成分包括：雀麦花叶病毒RNA、小麦胚无细胞蛋白质合成系统以及从小麦胚中提取的纯酪氨酸-tRNA合成酶。在用酪氨酸化和未酪氨酸化的雀麦花叶病毒RNA指导的体外蛋白质合成过程中，所形成的多肽的数量、种类以及翻译反应的速度均未观察到差异。在体外翻译过程中加入过量的活性酪氨酸-tRNA合成酶，翻译效率并未改变。在翻译之前，将雀麦花叶病毒RNA与活性酪氨酸-tRNA合成酶预先孵育，以便在没有翻译系统成分的情况下相互作用，然后进行体外翻译。当雀麦花叶病毒RNA与无活性的酪氨酸-tRNA合成酶或牛血清白蛋白预先孵育时，也得到了类似的结果。这些结果表明，雀麦花叶病毒RNA的氨酰化作用对体外病毒蛋白质合成没有显著影响。在雀麦花叶病毒RNA衣壳化过程中，酪氨酸化或未酪氨酸化的雀麦花叶病毒RNA 4都可以以类似的方式被衣壳化。
