AFFINITIES OF RIBOSOMAL-PROTEIN S20 AND C-TERMINAL DELETION MUTANTS FOR 16S RIBOSOMAL-RNA AND S20 MESSENGER-RNA
AFFINITIES OF RIBOSOMAL-PROTEIN S20 AND C-TERMINAL DELETION MUTANTS FOR 16S RIBOSOMAL-RNA AND S20 MESSENGER-RNA
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DOI：
10.1093/nar/16.3.997
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发表时间：
1988-02-11
影响因子：
14.9
通讯作者：
MACKIE, GA
中科院分区：
文献类型：
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作者：
DONLY, BC;MACKIE, GA
我们测量了大肠杆菌核糖体蛋白S20和一些c端缺失突变体与16S rRNA的结合，并在体外转录了S20 mRNA。利用合适的质粒模板，通过偶联转录和翻译，在体外合成了感兴趣的突变体s20。体外产生的S20对16S rRNA的亲和力为1.2倍。107 （M-1）的凝胶过滤试验。只要从S20的C端去除6个残基，就会导致结合活性的急剧丧失，这表明该区域存在关键残基。对S20的氨基酸序列分析表明，这些残基可能是. α的一部分。螺旋。虽然已知S20可以自动调节其自身的合成，但我们无法证明S20对其自身mRNA的任何可测量的亲和力。
We have measured the binding of Escherichia coli ribosomal protein S20 and a number of C-terminal deletion mutants to 16S rRNA and in vitro transcribed S20 mRNA. Mutant S20s of interest were synthesized in vitro from the appropriate plasmid templates by coupled transcription and translation. The affinity of S20 produced in vitro for 16S rRNA is 1.2 .times. 107 (M-1) in a gel filtration assay. Removal of as few as 6 residues from the C terminus of S20 results in a sharp loss of binding activity, suggesting the presence of critical residues in this region. Analysis of the amino acid sequence of S20 indicates that these residues may constitute part of a segment of .alpha. helix. Although S20 is known to autoregulate its own synthesis, we were unable to demonstrate any measurable affinity of S20 for its own mRNA.