Covalent complex of phenylalanyl‐tRNA synthetase with 4‐thiouridine‐substituted tRNAPhe gene transcript retains aminoacylation activity

FEBS Letters

Favre, Alain¹  Moor, Nina A²  Lavrik, Olga I² 

[1] Institut Jacques Monod, CNRS-Université Paris 7, 75251 Paris Cedex 05, France;Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of the Russian Academy of Sciences, Novosibirsk 630090, Russia

关键词: Protein-nucleic acid recognition;    Phenylalanyl-tRNA synthetase;    Affinity labeling;    4-Thiouridine;    Thermus thermophilus;    aaRS(s);    aminoacyl-tRNA synthetase(s);    E.C. 6.1.1;    APM;    (N-acryloylamino)phenylmercuric chloride;    PAG(E);    polyacrylamide gel (electrophoresis);    PheRS;    phenylalanyl-tRNA synthetase;    s4U;    4-thiouridine;    [1s4U]-tRNAPhe and [16s4U]-tRNAPhe;    tRNAPhe gene transcripts containing 1 or 16 s4U residues;    tRNAPhe(-pA76);    tRNAPhe lacking the 3′-terminal nucleotide;   

DOI  :  10.1016/S0014-5793(98)00398-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

s⁴U-containing transcripts of tRNA^Phe gene have been prepared by complete substitution of 16 U residues or by random incorporation of s⁴U residues followed by affinity electrophoresis isolation of s⁴U-monosubstituted tRNA transcripts. Both analogs have been cross-linked to Thermus thermophilus phenylalanyl-tRNA synthetase (PheRS) and the specificity of the cross-linking has been demonstrated. Functional activity of the covalent complex of PheRS with the s⁴U-monosubstituted transcript has been shown by aminoacylation of 60% of the enzyme-cross-linked tRNA. This is the first instance in which biological activity of aminoacyl-tRNA synthetase and cross-linked tRNA in a specific complex has been revealed.

【 授权许可】

Unknown