Mapping functional regions of transcription factor TFIIIA
Mapping functional regions of transcription factor TFIIIA
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转录因子 TFIIIA 的功能区域图谱
DOI：
--
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发表时间：
1988
影响因子：
5.3
通讯作者：
D D Brown
中科院分区：
文献类型：
--
作者：
K E Vrana;M E Churchill;T. Tullius;D D Brown
反式作用因子TFIIIA的功能缺失突变体，在分子的两端截短，通过体外转录cDNA克隆和随后合成的mRNAs的无细胞翻译来表达。TFIIIA羧基末端附近的19个氨基酸或更少的区域是最大转录的关键，位于DNA结合域之外。延长的蛋白质可以排列在非洲爪哇5S RNA基因的内部控制区(ICR)上，其羧基末端朝向基因的5‘端，氨基末端朝向基因的3’端。这9个“锌指”和连接它们的连接子构成了蛋白质质量的80%，对应于TFIIIA的DNA结合域。蛋白质氨基末端附近的锌指比羧基末端附近的锌指对蛋白质与ICR的总结合能贡献更大。TFIIIA最显著的特点是它的模块化结构。每个锌指只与ICR内三个短核苷酸序列中的一个结合，这证明了这一点。
Functional deletion mutants of the trans-acting factor TFIIIA, truncated at both ends of the molecule, have been expressed by in vitro transcription of a cDNA clone and subsequent cell-free translation of the synthetic mRNAs. A region of TFIIIA 19 amino acids or less, near the carboxyl terminus, is critical for maximal transcription and lies outside the DNA-binding domain. The elongated protein can be aligned over the internal control region (ICR) of the Xenopus 5S RNA gene with its carboxyl terminus oriented toward the 5' end of the gene and its amino terminus oriented toward the 3' end of the gene. The nine "zinc fingers" and the linkers that separate them comprise 80% of the protein mass and correspond to the DNA-binding domain of TFIIIA. The zinc fingers near the amino terminus of the protein contribute more to the overall binding energy of the protein to the ICR than do the zinc fingers near the carboxyl end. The most striking feature of TFIIIA is its modular structure. This is demonstrated by the fact that each zinc finger binds to just one of three short nucleotide sequences within the ICR.
羟基自由基足迹法：一种绘制蛋白质-DNA 接触图谱的高分辨率方法。
影响因子：
--
作者：
Tullius,TD;Dombroski,BA;Churchill,ME;Kam,L
通讯作者：
Kam,L
DOI：
10.1073/pnas.83.15.5469
发表时间：
1986-08-01
影响因子：
11.1
作者：
TULLIUS, TD;DOMBROSKI, BA
通讯作者：
DOMBROSKI, BA