AIMP3/p18 Controls Translational Initiation by Mediating the Delivery of Charged Initiator tRNA to Initiation Complex
Aminoacyl-tRNA synthetase-interacting multifunctional proteins (AIMPs) are nonenzymatic scaffolding proteins that comprise multisynthetase complex (MSC) with nine aminoacyl-tRNA synthetases in higher eukaryotes. Among the three AIMPs, AIMP3/p18 is strongly anchored to methionyl-tRNA synthetase (MRS) in the MSC. MRS attaches methionine (Met) to initiator tRNA (tRNAiMet) and plays an important role in translation initiation. It is known that AIMP3 is dispatched to nucleus or nuclear membrane to induce DNA damage response or senescence; however, the role of AIMP3 in translation as a component of MSC and the meaning of its interaction with MRS are still unclear. Herein, we observed that AIMP3 specifically interacted with Met-tRNAiMetin vitro, while it showed little or reduced interaction with unacylated or lysine-charged tRNAiMet. In addition, AIMP3 discriminates Met-tRNAiMet from Met-charged elongator tRNA based on filter-binding assay. Pullâdown assay revealed that AIMP3 and MRS had noncompetitive interaction with eukaryotic initiation factor 2 (eIF2) Î³ subunit (eIF2Î³), which is in charge of binding with Met-tRNAiMet for the delivery of Met-tRNAiMet to ribosome. AIMP3 recruited active eIF2Î³ to the MRS–AIMP3 complex, and the level of Met-tRNAiMet bound to eIF2 complex was reduced by AIMP3 knockdown resulting in reduced protein synthesis. All these results suggested the novel function of AIMP3 as a critical mediator of Met-tRNAiMet transfer from MRS to eIF2 complex for the accurate and efficient translation initiation. âº AIMP3 interacts with MRS in the MSC. âº AIMP3 specifically binds to Met-tRNAiMet, which is produced by MRS. âº AIMP3 recruits active eIF2Î³ to the MRS–AIMP3 complex. âº Depletion of AIMP3 reduces the formation of ternary complex and global translation. âº AIMP3 plays an important role in translation initiation via Met-tRNAiMet delivery.