Arc1p organizes the yeast aminoacyl-tRNA synthetase complex and stabilizes its interaction with the cognate tRNAs.
Eukaryotic aminoacyl-tRNA synthetases, in contrast to their prokaryotic counterparts, are often part of high molecular weight complexes. In yeast, two enzymes, the methionyl- and glutamyl-tRNA synthetases associate in vivo with the tRNA-binding protein Arc1p. To study the assembly and function of this complex, we have reconstituted it in vitro from individually purified recombinant proteins. Our results show that Arc1p can readily bind to either or both of the two enzymes, mediating the formation of the respective binary or ternary complexes. Under competition conditions, Arc1p alone exhibits broad specificity and interacts with a defined set of tRNA species. Nevertheless, the in vitro reconstituted Arc1p-containing enzyme complexes can bind only to their cognate tRNAs and tighter than the corresponding monomeric enzymes. These results demonstrate that the organization of aminoacyl-tRNA synthetases with general tRNA-binding proteins into multimeric complexes can stimulate their catalytic efficiency and, therefore, offer a significant advantage to the eukaryotic cell.
Deinert
K
K
Biochemie-Zentrum Heidelberg (BZH), University of Heidelberg, Im Neuenheimer Feld 328, D-69120 Heidelberg, Germany.
Fasiolo
F
F
Hurt
E C
EC
Simos
G
G
D002384:Catalysis; D005057:Eukaryotic Cells / Q000201:enzymology; D019143:Evolution, Molecular; D005975:Glutamate-tRNA Ligase / Q000378:metabolism*; D046911:Macromolecular Substances; D008718:Methionine-tRNA Ligase / Q000378:metabolism*; D008958:Models, Molecular; D010441:Peptide Chain Elongation, Translational; D011485:Protein Binding; D011487:Protein Conformation; D012343:RNA, Transfer / Q000378:metabolism*; D012352:RNA, Transfer, Glu / Q000378:metabolism; D012358:RNA, Transfer, Met / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D013379:Substrate Specificity; D015003:Yeasts
C102145:ARC1 protein, S cerevisiae; D046911:Macromolecular Substances; D012352:RNA, Transfer, Glu; D012358:RNA, Transfer, Met; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D012343:RNA, Transfer; D008718:Methionine-tRNA Ligase; D005975:Glutamate-tRNA Ligase