Escherichia coli proline tRNA: structure and recognition sites for prolyl-tRNA synthetase.

T Hasegawa, T Yokogawa
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引用次数: 10

Abstract

A major proline tRNA was purified from bulk Escherichia coli A19 tRNA by affinity chromatography with a biotinylated DNA probe. Its nucleotide sequence including modified nucleotides was determined by the post-labelling technique. In order to study the recognition sites of this proline tRNA for prolyl-tRNA synthetase, various mutant transcripts were prepared using an in vitro transcription system with T7 RNA polymerase. Based on the results of in vitro kinetic analyses of mutant transcripts, it was concluded that the second and third letters, G35 and G36, of the anticodon, G37 of the anticodon loop, the discriminator base A73, G72 of the acceptor stem, G49 and U17A that existed in the corner of an L-shaped structure are the recognition sites of proline tRNA for prolyl-tRNA synthetase.

大肠杆菌脯氨酸tRNA:脯氨酸tRNA合成酶的结构和识别位点。
用生物素化DNA探针亲和层析法从大肠杆菌A19 tRNA中纯化了一个主要的脯氨酸tRNA。通过后标记技术确定其核苷酸序列,包括修饰的核苷酸。为了研究脯氨酸tRNA对脯氨酸-tRNA合成酶的识别位点,利用T7 RNA聚合酶体外转录系统制备了多种突变体转录本。根据突变体转录本的体外动力学分析结果,认为反密码子的第2和第3个字母G35和G36、反密码子环的G37、鉴别碱基A73、受体茎的G72、l型结构角上的G49和U17A是脯氨酸tRNA对脯氨酸tRNA合成酶的识别位点。
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