Purification of active RNA polymerase I from yeast.
Purification of active RNA polymerase I from yeast.
复制标题
从酵母中纯化活性 RNA 聚合酶 I。
DOI：
10.1007/978-1-4939-2392-2_16
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Schneider,DavidAlan
中科院分区：
文献类型：
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作者：
Appling,FrancisDean;Schneider,DavidAlan
真核细胞使用至少三种核DNA依赖性RNA聚合酶系统来合成细胞RNA。RNA聚合酶I、II和III主要分别产生rRNA、mRNA和tRNA。在快速生长的细胞中，大部分RNA合成都致力于翻译机器的产生，其中RNA聚合酶I的rRNA合成占总细胞转录的一半以上。核糖体生物发生和细胞生长之间的基本联系是明确的;此外，最近的研究已经确定RNA聚合酶I的转录是抗癌化疗的关键靶点。因此，需要用于表征核糖体DNA的转录及其调节的有效方法。为了描述酶的酶特性，体外测定是关键的。在这里，我们描述了一种纯化RNA聚合酶I的方法。这种方法产生足够高的数量和活性的酶，用于一系列旨在详细描述RNA聚合酶I的酶性质的实验。
Eukaryotic cells employ at least three nuclear, DNA-dependent RNA polymerase systems for the synthesis of cellular RNA. RNA polymerases I, II, and III primarily produce rRNA, mRNA, and tRNA, respectively. In a rapidly growing cell, most RNA synthesis is devoted to production of the translation machinery, with rRNA synthesis by RNA polymerase I representing more than half of total cellular transcription. The fundamental connection between ribosome biogenesis and cell growth is clear; furthermore, recent studies have identified transcription by RNA polymerase I as a key target for anticancer chemotherapy. Thus, efficient methods for characterizing transcription of the ribosomal DNA and its regulation are needed. In order to describe enzymatic features of an enzyme, in vitro assays are critical. Here we describe a method for purifying RNA polymerase I. This approach yields enzyme of sufficiently high quantity and activity for an array of experiments directed at describing the enzymatic properties of RNA polymerase I in detail.