REGIONAL CODON RANDOMIZATION - DEFINING A TATA-BINDING PROTEIN SURFACE REQUIRED FOR RNA POLYMERASE-III TRANSCRIPTION
REGIONAL CODON RANDOMIZATION - DEFINING A TATA-BINDING PROTEIN SURFACE REQUIRED FOR RNA POLYMERASE-III TRANSCRIPTION
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DOI：
10.1126/science.8211143
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发表时间：
1993-10-08
期刊：
影响因子：
56.9
通讯作者：
STRUHL, K
中科院分区：
文献类型：
--
作者：
CORMACK, BP;STRUHL, K
TATA结合蛋白(TBP)是三种核RNA聚合酶转录所必需的。TBP受到区域密码子随机化的影响，这是一种基于密码子的突变方法，可以产生复杂而紧凑的蛋白质文库。对186个温度敏感型TBP突变体的分析发现，有65个突变在RNA聚合酶III(POL III)的转录中存在特异性缺陷。这些突变体映射到一个有限的TBP表面，该表面可能与POLIII转录因子TFIIIB的一个组成部分Tds4相互作用。含有Pol III缺陷衍生物的菌株的信使RNA数量增加，这表明TBP相互作用因子之间对限制TBP数量的竞争决定了体内Pol II和Pol III转录的比率。
The TATA-binding protein (TBP) is required for transcription by all three nuclear RNA polymerases. TBP was subjected to regional codon randomization, a codon-based mutagenesis method that generates complex yet compact protein libraries. Analysis of 186 temperature-sensitive TBP mutants yielded 65 specifically defective in transcription by RNA polymerase III (Pol III). These mutants map to a limited TBP surface that may interact with Tds4, a component of the Pol III transcription factor TFIIIB. Strains that contain the Pol III-defective derivatives have increased amounts of messenger RNA, which suggests that competition among TBP-interacting factors for limiting quantities of TBP determines the ratio of Pol II and Pol III transcription in vivo.