Functional interplay between arginyl-tRNA synthetases and arginyltransferase

Abstract

Protein arginylation, mediated by arginyltransferase ATE1, is a posttranslational modification of emerging biological importance that consists of transfer of the amino acid Arg to protein and peptide substrates. ATE1 utilizes charged tRNAArg as the donor of the arginyl group, which depends on the activity of Arg-tRNA synthetases (RARS) and is also utilized in translation. The mechanisms that regulate the functional balance between ATE1, RARS and translation are unknown. Here we addressed the functional interplay between these mechanisms using intracellular arginylation sensor in cell lines with overexpression or deletion of ATE1 and RARS isoforms. We find that arginylation levels depend on the physiological state of the cells but are not directly affected by translation activity or availability of RARS isoforms. However, displacement of RARS from the multi-synthetase complex leads to an increase in intracellular arginylation independently of RARS enzymatic activity. This effect is accompanied by ATE1s redistribution into the cytosol. Our results provide the first comprehensive analysis of the interdependence between translation, arginyl-tRNA synthesis, and arginylation. Key PointsO_LIIntracellular arginylation depends on the physiological state of the cell, but does not compete with the translation machinery C_LIO_LIA fraction of ATE1 binds directly to both long and short Arg-tRNA synthetases (RARS) C_LIO_LIDisplacement of long RARS from the multi-tRNA synthetase complex increases cytosolic fraction and activity of ATE1 C_LI