title: Reinvestigation of aminoacyl-tRNA synthetase core complex by affinity purification-mass spectrometry reveals TARSL2 as a potential member of the complex.
abstract: Twenty different aminoacyl-tRNA synthetases (ARSs) link each amino acid to their cognate tRNAs. Individual ARSs are also associated with various non-canonical activities involved in neuronal diseases, cancer and autoimmune diseases. Among them, eight ARSs (D, EP, I, K, L, M, Q and RARS), together with three ARS-interacting multifunctional proteins (AIMPs), are currently known to assemble the multi-synthetase complex (MSC). However, the cellular function and global topology of MSC remain unclear. In order to understand the complex interaction within MSC, we conducted affinity purification-mass spectrometry (AP-MS) using each of AIMP1, AIMP2 and KARS as a bait protein. Mass spectrometric data were funneled into SAINT software to distinguish true interactions from background contaminants. A total of 40, 134, 101 proteins in each bait scored over 0.9 of SAINT probability in HEK 293T cells. Complex-forming ARSs, such as DARS, EPRS, IARS, Kars, LARS, MARS, QARS and RARS, were constantly found to interact with each bait. Variants such as, AIMP2-DX2 and AIMP1 isoform 2 were found with specific peptides in KARS precipitates. Relative enrichment analysis of the mass spectrometric data demonstrated that TARSL2 (threonyl-tRNA synthetase like-2) was highly enriched with the ARS-core complex. The interaction was further confirmed by coimmunoprecipitation of TARSL2 with other ARS core-complex components. We suggest TARSL2 as a new component of ARS core-complex.
authors: Biomedical Research Institute, Korea Institute of Science and Technology, Seongbuk-gu, Seoul, Korea ; School of Life Sciences and Biotechnology, Korea University, Seongbuk-gu, Seoul, Korea.
authors: Kyutae
authors: K
authors: Kim
authors: Seong-Jun
authors: SJ
authors: Park
authors: Seungjin
authors: S
authors: Na
authors: Jun Seok
authors: JS
authors: Kim
authors: Hyungwon
authors: H
authors: Choi
authors: Yoon Ki
authors: YK
authors: Kim
authors: Eunok
authors: E
authors: Paek
authors: Cheolju
authors: C
authors: Lee
mesh_terms: D000465:Algorithms; D000595:Amino Acid Sequence; D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry* / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism; D002846:Chromatography, Affinity; D019295:Computational Biology / Q000379:methods*; D016207:Cytokines / Q000737:chemistry / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008250:Lysine-tRNA Ligase / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism; D009687:Nuclear Proteins; D025941:Protein Interaction Mapping / Q000379:methods*; D011499:Protein Processing, Post-Translational; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D013914:Threonine-tRNA Ligase / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: C575685:AIMP2 protein, human; D002352:Carrier Proteins; D016207:Cytokines; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; C077219:small inducible cytokine subfamily E, member 1; D000604:Amino Acyl-tRNA Synthetases; C000591800:TARS3 protein, human; D013914:Threonine-tRNA Ligase; D008250:Lysine-tRNA Ligase