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Cell‐based analysis of pairwise interactions between the components of the multi‐tRNA synthetase complex
Author(s) -
Kong Jiwon,
Kim Sunghoon
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.202000418r
Subject(s) - aminoacyl trna synthetase , transfer rna , biology , ribosome , protein biosynthesis , microbiology and biotechnology , translation (biology) , biochemistry , complementation , cytoplasm , rna , gene , messenger rna , phenotype
Cytoplasmic aminoacyl‐tRNA synthetases (ARSs) are organized into multi‐tRNA synthetase complexes (MSCs), from Archaea to mammals. An evolutionary conserved role of the MSCs is enhancement of aminoacylation by forming stable associations of the ARSs and tRNAs. In mammals, a single macromolecular MSC exists, which is composed of eight cytoplasmic ARSs, for nine amino acids, and three scaffold proteins. Consequently, nearly half of aminoacyl‐tRNA efflux becomes concentrated at the MSC. Stable supply of aminoacyl‐tRNA to the ribosome is, therefore, considered to be a major role of the mammalian MSC. Furthermore, the mammalian MSC also serves as a reservoir for releasable components with noncanonical functions. In this study, a split‐luciferase complementation system was applied to investigate the configuration of the MSC in live mammalian cells. Multiplex interconnections between the components were simplified into binary protein‐protein interactions, and pairwise comparison of the interactions reconstituted a framework consistent with previous in vitro studies. Reversibility of the split‐luciferase reporter binding demonstrated convertible organization of the mammalian MSC, including interferon gamma (IFNγ)‐stimulated glutamyl‐prolyl‐tRNA synthetase 1 (EPRS1) release, as well as the cooperation with the ribosome bridged by the tRNAs. The cell‐based analysis provided an improved understanding of the flexible framework of the mammalian MSC in physiological conditions.