Random‐splitting of tRNA transcripts as an approach for studying tRNA‐protein interactions

FEBS Letters

Kisselev, Lev¹  Pugachev, Vladimir¹  Beresten, Sergey¹  Aphasizhev, Ruslan¹ 

[1] Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 117984, Russian Federation

关键词: RNA-transcripts;    Beef;    Tryptophanyl-tRNA synthetase;    Aminoacylation;    b (prefix);    bovine;    QRS;    glutaminyl-tRNA synthetase;    WRS;    tryptophanyl-tRNA synthetase;   

DOI  :  10.1016/0014-5793(93)81474-E

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Location of phosphodiester bonds essential for aminoacylation of bovine tRN^Trp was identified using a randomly cleaved transcript synthesized in vitro. It was found that cleavage of phosphodiester bonds after nucleotides in positions 21, 22, 36–38, 57–59, 62 and 64 were critical for aminoacylation capacity of tRNA^Trp-transcript. These cleavage sites were located in the regions of tRNA molecule protected by the cognate synthetase against chemical modification and in the regions presumably outside the contact area as well. These results indicate that for maintenance of aminoacylation ability the intactness of the certain regions of the tRNA backbone structure is necessary. Random splitting of non-modified RNA with alkali followed by separation of active and inactive molecules and identification of cleavage sites developed in this work may become a general approach for studying the role of RNA covalent structure in its interaction with proteins.

【 授权许可】

Unknown