A new method for identifying the amino acid attached to a particular RNA in the cell.
To investigate the function of tRNAs or any other aminoacylable RNAs in vivo, it is important to be able to estimate the amounts and species of aminoacylated RNAs in living cells. We have developed a method of analyzing amino acids attached to particular tRNAs obtained from cells. After the ester bond between the amino acid and the 3'-adenosine moiety of a specific aminoacyl-tRNA is stabilized by acetylation of the amino acid with [14C]acetic anhydride, the aminoacyl-tRNA can be fished out with a solid-phase-attached DNA probe. The 14C-labeled acetylamino acid is then released from the thus purified acetyl-aminoacyl-tRNAs by alkaline treatment and detected by TLC analysis.
Suzuki
T
T
Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo, Japan.
Ueda
T
T
Watanabe
K
K
D000089:Acetic Anhydrides; D000107:Acetylation; D000596:Amino Acids / Q000032:analysis*; D001483:Base Sequence; D002175:Candida / Q000378:metabolism; D002250:Carbon Radioisotopes; D002855:Chromatography, Thin Layer; D015342:DNA Probes; D007202:Indicators and Reagents; D007700:Kinetics; D007930:Leucine / Q000031:analogs & derivatives / Q000032:analysis; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012331:RNA, Fungal / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D012694:Serine / Q000031:analogs & derivatives / Q000032:analysis
D000089:Acetic Anhydrides; D000596:Amino Acids; D002250:Carbon Radioisotopes; D015342:DNA Probes; D007202:Indicators and Reagents; D015345:Oligonucleotide Probes; D012331:RNA, Fungal; D012346:RNA, Transfer, Amino Acyl; C031800:acetic anhydride; D012694:Serine; D007930:Leucine; C088117:acetylleucine; C012162:N-acetylserine