Greider C W, Blackburn E H
Department of Molecular Biology, University of California, Berkeley 94720.
Cell. 1987 Dec 24;51(6):887-98. doi: 10.1016/0092-8674(87)90576-9.
We have analyzed the de novo telomere synthesis catalyzed by the enzyme telomere terminal transferase (telomerase) from Tetrahymena. Oligonucleotides representing the G-rich strand of telomeric sequences from five different organisms specifically primed the addition of TTGGGG repeats in vitro, suggesting that primer recognition may involve a DNA structure unique to these oligonucleotides. The sequence at the 3' end of the oligonucleotide primer specified the first nucleotide added in the reaction. Furthermore, the telomerase was shown to be a ribonucleoprotein complex whose RNA and protein components were both essential for activity. After extensive purification of the enzyme by a series of five different chromatographic steps, a few small low abundance RNAs copurified with the activity.
我们分析了来自嗜热四膜虫的端粒末端转移酶(端粒酶)催化的从头端粒合成。代表五种不同生物体端粒序列富含G链的寡核苷酸在体外特异性引发了TTGGGG重复序列的添加,这表明引物识别可能涉及这些寡核苷酸特有的DNA结构。寡核苷酸引物3'端的序列决定了反应中添加的第一个核苷酸。此外,端粒酶被证明是一种核糖核蛋白复合物,其RNA和蛋白质成分对活性都是必不可少的。通过一系列五个不同的色谱步骤对该酶进行广泛纯化后,一些少量低丰度RNA与活性一起共纯化。