Institut für Biochemie, Universität Würzburg, Röntgenring 11, D-8700 Würzburg, FRG.
EMBO J. 1983;2(4):605-10. doi: 10.1002/j.1460-2075.1983.tb01470.x.
tRNA preparations from Chlamydomonas and wheat germ contain small amounts of tRNA 5' halves and corresponding 3' halves. Incubation of cell-free extracts from the two sources with [gamma-P]ATP yielded 5'-P-labeled tRNA 3' halves which were joined to their corresponding 5' counterparts to form mature tRNA containing 2'-phosphomonoester,3', 5'-phosphodiester bonds. tRNA 3' halves labelled with T4 kinase were purified, sequenced and also joined to their 5' counterparts. It is proposed that these tRNA halves may be intermediates of the tRNA splicing process, and that the RNA kinase and ligase activities observed here are part of the tRNA splicing complex.
来自衣藻和麦胚的 tRNA 制剂含有少量的 tRNA 5' 片段和相应的 3' 片段。用 [γ-P]ATP 孵育这两种来源的无细胞提取物,得到了 5'-P 标记的 tRNA 3' 片段,这些片段与相应的 5' 片段连接,形成含有 2'-磷酸单酯、3'、5'-磷酸二酯键的成熟 tRNA。用 T4 激酶标记的 tRNA 3' 片段被纯化、测序,并与它们的 5' 片段连接。据推测,这些 tRNA 片段可能是 tRNA 剪接过程的中间产物,并且这里观察到的 RNA 激酶和连接酶活性是 tRNA 剪接复合物的一部分。