Construction, aminoacylation and 80 S ribosomal complex formation with a yeast initiator tRNA having an arginine CCU anticodon

FEBS Letters

Grosjean, Henri¹  Beauchemin, Nicole¹  Cedergren, Robert¹ 

[1] Département de Biochimie, Université de Montréal, C.P. 6128, Montréal, H3C 3J7, Canada

关键词: initiator tRNA recombinant RNA Aminoacylation Ribosomal complex;   

DOI  :  10.1016/0014-5793(86)80639-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The digestion of yeast initiator methionine tRNA with mung bean nuclease and U₂ ribonuclease yielded 5'- and 3'-fragments, respectively. These two fragments together represent the entire tRNA sequence except for A₃₅, the central nucleotide of the anticodon, and the CCA terminus. Using RNA ligase, a cytosine was added and the anticodon loop having a C₃₅ was reformed. Subsequent treatment of this product with CCA-transferase yielded a full-length methionine tRNA having an arginine CCU anticodon. This recombinant tRNA^Met (CCU) was charged with methionine by the yeast tRNA synthetase. Aminoacylation of the recombinant was however less extensive than in the case of native tRNA^Met (CAU). After aminoacylation the recombinant tRNA formed an 80 S ribosomal complex.

【 授权许可】

Unknown