The RNA sequence context defines the mechanistic routes by which yeast arginyl-tRNA synthetase charges tRNA.
Arginylation of tRNA transcripts by yeast arginyl-tRNA synthetase can be triggered by two alternate recognition sets in anticodon loops: C35 and U36 or G36 in tRNA(Arg) and C36 and G37 in tRNA(Asp) (Sissler M, Giegé R, Florentz C, 1996, EMBO J 15:5069-5076). Kinetic studies on tRNA variants were done to explore the mechanisms by which these sets are expressed. Although the synthetase interacts in a similar manner with tRNA(Arg) and tRNA(Asp), the details of the interaction patterns are idiosyncratic, especially in anticodon loops (Sissler M, Eriani G, Martin F, Giegé R, Florentz C, 1997, Nucleic Acids Res 25:4899-4906). Exchange of individual recognition elements between arginine and aspartate tRNA frameworks strongly blocks arginylation of the mutated tRNAs, whereas full exchange of the recognition sets leads to efficient arginine acceptance of the transplanted tRNAs. Unpredictably, the similar catalytic efficiencies of native and transplanted tRNAs originate from different k(cat) and Km combinations. A closer analysis reveals that efficient arginylation results from strong anticooperative effects between individual recognition elements. Nonrecognition nucleotides as well as the tRNA architecture are additional factors that tune efficiency. Altogether, arginyl-tRNA synthetase is able to utilize different context-dependent mechanistic routes to be activated. This confers biological advantages to the arginine aminoacylation system and sheds light on its evolutionary relationship with the aspartate system.
Sissler
M
M
UPR 9002, IBMC du CNRS, Strasbourg, France.
Giegé
R
R
Florentz
C
C
D000926:Anticodon; D001126:Arginine-tRNA Ligase / Q000378:metabolism*; D001483:Base Sequence; D019143:Evolution, Molecular; D005656:Fungal Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D014176:Protein Biosynthesis; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012347:RNA, Transfer, Arg / Q000235:genetics / Q000378:metabolism*; D012349:RNA, Transfer, Asp / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D015003:Yeasts
D000926:Anticodon; D005656:Fungal Proteins; D012331:RNA, Fungal; D012347:RNA, Transfer, Arg; D012349:RNA, Transfer, Asp; D001126:Arginine-tRNA Ligase