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首页> 外文期刊>Journal of Molecular Biology >The role of the central zinc fingers of transcription factor IIIA in binding to 5 S RNA.
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The role of the central zinc fingers of transcription factor IIIA in binding to 5 S RNA.

机译：转录因子IIIA的中央锌指在与5 S RNA结合中的作用。

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摘要

In the nine-zinc finger Xenopus transcription factor TFIIIA the central group of fingers, fingers 4 to 7, have been shown to bind to 5 S RNA. In this study, we have attempted to assess the role of this region of the TFIIIA molecule in more detail than hitherto. High-resolution footprinting by RNases A and CV1 has been used to probe the binding to 5 S RNA of three TFIIIA peptides Tf(1-6), Tf(4-6) and Tf(4-7), consisting of fingers 1 to 6, 4 to 6, and 4 to 7, respectively, and of full-length TFIIIA. The results pinpoint the outer margins of binding of the central fingers within helices IV and II of TFIIIA. A comparison of the footprints reveals that the presence of finger 7 affords protection at positions C19 and U55, the distal portion of helix II and the proximal portion of loop B. In addition, our footprints suggest that the central fingers bind in the same manner, whether in an isolated group or in the intact TFIIIA molecule. In a companion study, we have determined the binding affinities of Tf(4-6) and Tf(4-7) for full-length and three truncated 5 S RNA molecules, the latter selected on the basis of the regions of the 5 S RNA molecule known to be important in the binding of TFIIIA. The analysis uses only fully active protein involved in the binding and the results are consistent with the corresponding footprinting results. This is the first time that a detailed study of the binding site of one particular zinc finger to RNA has been reported; the results should be of use in the design of 5 S RNA molecules and TFIIIA peptides for structural studies of the interaction between zinc fingers and RNA. Copyright 2000 Academic Press.
机译：在九锌手指非洲爪蟾转录因子TFIIIA中，中心手指（手指4至7）已显示与5 S RNA结合。在这项研究中，我们试图比迄今为止更详细地评估TFIIIA分子这一区域的作用。 RNases A和CV1的高分辨率足迹已用于探测与3个TFIIIA肽Tf（1-6），Tf（4-6）和Tf（4-7）的5 S RNA的结合，由手指1至全长TFIIIA分别为6、4至6和4至7。结果精确指出了TFIIIA螺旋IV和II中中心指结合的外部边缘。足迹比较表明，手指7的存在在位置C19和U55，螺旋II的远端和环B的近端提供了保护。此外，我们的足迹表明，中央手指以相同的方式绑定，无论是孤立的组还是完整的TFIIIA分子。在同伴研究中，我们确定了Tf（4-6）和Tf（4-7）对全长和三个截短的5S RNA分子的结合亲和力，后者是根据5S区域选择的已知在TFIIIA的结合中起重要作用的RNA分子。该分析仅使用参与结合的完全活性蛋白，其结果与相应的印迹结果一致。这是首次报道了对一个特定锌指与RNA结合位点的详细研究。该结果应可用于设计5 S RNA分子和TFIIIA肽，用于锌指与RNA之间相互作用的结构研究。版权所有2000学术出版社。

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