Nuclease protection analysis of ribonucleoprotein complexes: use of the cytotoxic ribonuclease alpha-sarcin to determine the binding sites for Escherichia coli ribosomal proteins L5, L18, and L25 on 5S rRNA.
Nuclease protection analysis of ribonucleoprotein complexes: use of the cytotoxic ribonuclease alpha-sarcin to determine the binding sites for Escherichia coli ribosomal proteins L5, L18, and L25 on 5S rRNA.
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核糖核蛋白复合物的核酸酶保护分析：使用细胞毒性核糖核酸酶 α-sarcin 确定大肠杆菌核糖体蛋白 L5、L18 和 L25 在 5S rRNA 上的结合位点。
DOI：
10.1073/pnas.81.2.322
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发表时间：
1984
影响因子：
11.1
通讯作者：
Wool,IG
中科院分区：
文献类型：
--
作者：
Huber,PW;Wool,IG
设计了一种快速简便的方法来确定RNA上蛋白质的结合位点。该程序是一个适应的一个用于映射DNA-蛋白质复合物的保护免受核酸酶消化。该方法使用细胞毒性核糖核酸酶α-八叠球菌素，其水解RNA的单链和双链区域中的嘌呤。通过确认大肠杆菌核糖体蛋白L18和L25在5S rRNA上的结合位点对其进行了验证，并通过鉴定蛋白L5的附着位点确定了其价值。该方法对其他核糖核蛋白复合物的分析也有一定的参考价值。
A rapid and convenient method has been devised to determine the binding sites for proteins on RNA. The procedure is an adaptation of one used to map DNA-protein complexes by protection against nuclease digestion. The method uses the cytotoxic ribonuclease alpha-sarcin, which hydrolyzes purines in both single- and double-stranded regions of RNA. It has been authenticated by confirming the binding sites for the Escherichia coli ribosomal proteins L18 and L25 on 5S rRNA and its value has been established by identifying the attachment site for protein L5. The procedure should be useful for the analysis of other ribonucleoprotein complexes.