Horikoshi M, Yamamoto T, Ohkuma Y, Weil P A, Roeder R G
Laboratory of Biochemistry and Molecular Biology, Rockefeller University, New York, New York 10021.
Cell. 1990 Jun 29;61(7):1171-8. doi: 10.1016/0092-8674(90)90681-4.
A systematic series of N-terminal, C-terminal, and internal deletion mutants of S. cerevisiae TFIID were expressed in vitro and tested for TATA box binding and basal level transcription activities using, respectively, DNA mobility shift and in vitro transcription assays. The domains responsible for these activities were colocalized to a surprisingly large region containing C-terminal residues 63-240. This region was noted previously to contain potentially interesting structural motifs (central basic core, direct repeats, and sigma factor homology) and, more recently, to be highly conserved among TFIID from different species. Deletion mutant cotranslation studies revealed that TFIID binds DNA as a monomer. The implications of these results for TFIID structure and function are discussed.
对酿酒酵母TFIID的一系列N端、C端和内部缺失突变体进行了体外表达，并分别使用DNA迁移率变动分析和体外转录分析测试了它们与TATA盒的结合能力和基础水平转录活性。负责这些活性的结构域共定位于一个惊人的大区域，该区域包含C端残基63 - 240。此前已注意到该区域含有潜在有趣的结构基序（中央碱性核心、直接重复序列和sigma因子同源性），最近还发现它在不同物种的TFIID中高度保守。缺失突变体共翻译研究表明，TFIID以单体形式结合DNA。讨论了这些结果对TFIID结构和功能的影响。
