Interaction of tRNAPhe and tRNAVal with Aminoacyl-tRNA Synthetases
A Chemical Modication Study
Article first published online: 3 MAR 2005
European Journal of Biochemistry
Volume 132, Issue 3, pages 537–544, May 1983
How to Cite
VLASSOV, V. V., KERN, D., ROMBY, P., GIEBÉ, R. and EBEL, J.-P. (1983), Interaction of tRNAPhe and tRNAVal with Aminoacyl-tRNA Synthetases. European Journal of Biochemistry, 132: 537–544. doi: 10.1111/j.1432-1033.1983.tb07395.x
- Issue published online: 3 MAR 2005
- Article first published online: 3 MAR 2005
- (Received November 25, 1982) – EJB 6256
The alkylation by ethylnitrosourea of phyosphodiester bonds in tRNAPhe from veast and in tRNAVal from yeast and from rabbit liver and that by 4-N-2-chloroethyl-n-methylamino)-benzylamine of N-7 atoms of guanosine residues in yeast tRNAVal have been used to study the interaction of these tRAAs with anminoacyl-tRNA synthetases. The modifications occurring at low yield were carried out on 3′ and'or 5′ end-labelled tRNAs either free or in the presence of cognate or non-cognate synthetases. After splitting of the tRNAs at the alkylated positions, the position of the modification sites in the tRNA sequences were detected by acrylamide gel electrophoresis. It was found that the synthetases protect against alkylation certain phosphate or guanosine residues in their cognate tRNAs. Non-cognate synthetases failed to protedct efficiently specific positions in tRNA against modification.
In yeast tRNAPhe the cognate phenylalanyl-tRNA synthetase protects certain phosphates located in all four stems and in the anticodon and extra-loop of the tRMA. Particularly strong protections occur on phosphate 34 in the anticodon loop and on phosphates 23, 27, 28, 41 in the D and anticodon stems.
In yeast tRNAVal complexed with yeast valyl-tRNA synthetase the protected phosphates are essentialy located in the tRNA. Three guanosine residues, located in the D stem, and another one in the 3′ part of the anticodon stem were also found protected by the synthetase. In mammalian tRNAVal, complexed with the congnate but heterologous yeasst valyl-tRNA synthetase, the protected phosphates lie in the anticodon stem, in the extra-loop andd in the T ψ arm.
The location of the protected residues in the structure of three tRNAs suggests some common features in the binding of tRNAs to aminoacyl-tRNA synthetases. These results will be discussed in the light of infsormations of interaction sites obtained by nucleases digestion and ultravilet cross-linking methods.