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Abstract
Aminoacyl-tRNA synthetases (aaRSs) are essential for decoding the genetic code by ligating amino acids to their cognate tRNAs with high specificity. This thesis investigates substrate recognition mechanisms in glutaminyl- and glutamyl-tRNA synthetases (GlnRS and GluRS), revealing nucleic acid recognition signatures within tRNA that influence both tRNA and amino acid specificity. Using rational engineering, we demonstrate that these signatures assist amino acid recognition, introducing a novel concept in genetic code translation. These findings provide generalized principles for protein–nucleic acid interactions and offer strategies for genetic code expansion in synthetic biology.