VALYL-TRNA SYNTHETASE INTERACTS WITH Β-SUBUNIT OF THE EUKARYOTIC TRANSLATION ELONGATION FACTOR COMPLEX eEF1B

Abstract

The aim of this study was to investigate the interaction of the N-terminal domain of the valyl-tRNA synthetase with α, β, and γ subunits of the eEF1B translation elongation factor complex. Methods: for this purpose, all 4 proteins were synthesized in bacterial cells and purified to homogeneity by a combination of chromatographic methods. The interaction of the eEF1B complex subunits with the N-terminal domain of the valyl-tRNA synthetase was verified by gel filtration and in vitro pull-down assays. Protein fractions collected at these stages were analyzed by SDS-PAGE. Results: according to the gel filtration results, eEF1Bα and eEF1Bγ subunits do not form a stable complex with the valine-tRNA synthetase domain. The potential for complexation of the eEF1Bβ subunit was evaluated by pull-down assay, which showed that this protein does interact with the valyl-tRNA synthetase. Conclusions: we concluded that the eEF1Bα and eEF1Bγ subunits do not interact with the valyl-tRNA synthetase compared to the eEF1Bβ protein. The N-terminal domain of the valyl-tRNA synthetase is necessary and sufficient for this interaction.