Aminoacylation of Synthetic DNAs Corresponding to Escherichia coli Phenylalanine and Lysine tRNAs

• Published in: Science (American Association for the Advancement of Science) 1988-07, Vol.241 (4861), p.74-79
• Main Authors: Khan, Akbar S., Roe, Bruce A.
• Format: Article
• Language: English
• Subjects: Amino acids; Aminoacyl-tRNA; Arsenates; Arsenic; Bacteria; Base Sequence; Biological and medical sciences; Deoxyuridine - metabolism; Dimethyl Sulfoxide - pharmacology; DNA; DNA - genetics; DNA - metabolism; Escherichia coli; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Genetics; Hydrogen-Ion Concentration; Kinetics; Lysine-tRNA Ligase - metabolism; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Nucleic acids; Phenylalanine-tRNA Ligase - metabolism; RNA; RNA, Bacterial - genetics; RNA, Transfer, Amino Acyl - genetics; Science; Sodium; Spermidine - pharmacology; Structure-Activity Relationship; Thymidine - metabolism; Transfer RNA; Translation. Translation factors. Protein processing
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.2455342

Synthetic DNA oligomers (tDNAs) corresponding to Escherichia coli tRNA$^{\text{Phe}}$ or tRNA$^{\text{Lys}}$ have been synthesized with either deoxythymidine (dT) or deoxyuridine (dU) substituted in the positions occupied by ribouridine or its derivatives. The tDNAs inhibited the aminoacylation of their respective tRNAs with their cognate amino acids, but not the aminoacylation of tRNA$^{\text{Leu}}$ with Leu. In the presence of aminoacyl-tRNA synthetase, species of both a tDNA$^{\text{Phe}}$ synthesized with a 3$^{\prime}$ terminal riboadenosine and a tDNA$^{\text{Lys}}$ containing only deoxynucleotides could be aminoacylated with the appropriate amino acids, although the Michaelis constant K$_{\text{m}}$ and observed maximal rate V$_{\max}$ values for aminoacylation were increased by three- to fourfold and decreased by two- to threefold, respectively. The aminoacylation of synthetic tDNAs demonstrates that the ribose backbone of a tRNA is not absolutely required for tRNA aminoacylation.