SEQUENCE-SPECIFIC INTERACTION OF R17-COAT PROTEIN WITH ITS RIBONUCLEIC-ACID BINDING-SITE
SEQUENCE-SPECIFIC INTERACTION OF R17-COAT PROTEIN WITH ITS RIBONUCLEIC-ACID BINDING-SITE
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DOI：
10.1021/bi00280a002
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发表时间：
1983-01-01
期刊：
影响因子：
2.9
通讯作者：
UHLENBECK, OC
中科院分区：
文献类型：
--
作者：
CAREY, J;CAMERON, V;UHLENBECK, OC
作为序列特异性RNA-蛋白质相互作用的一个例子，研究了噬菌体R17外壳蛋白与其RNA结合位点之间的相互作用，以进行翻译抑制。核酸酶保护和选择实验确定了与. apprx的结合位点。20个连续的核苷酸形成一个发夹。硝酸纤维素过滤器保留测定用于显示外壳蛋白和合成的21个核苷酸的RNA片段之间的结合符合简单的生物分子反应。单位化学计量和Kd为apprx。在2 ℃下获得1 nM。C在含有0.19 M盐的缓冲液中。这种相互作用是高度序列特异性的，因为多种RNA不能与21个核苷酸片段竞争外壳蛋白结合。
The interaction between phage R17 coat protein and its RNA binding site for translational repression was studied as an example of a sequence-specific RNA-protein interaction. Nuclease protection and selection experiments define the binding site to .apprx. 20 contiguous nucleotides which form a hairpin. A nitrocellulose filter retention assay is used to show that the binding between the coat protein and a synthetic 21 nucleotide RNA fragment conforms to a simple biomolecular reaction. Unit stoichiometry and a Kd of .apprx. 1 nM are obtained at 2.degree. C in buffer containing 0.19 M salt. The interaction is highly sequence specific since a variety of RNA failed to compete with the 21 nucleotide fragment for coat protein binding.