Purification and characterization of two serine isoacceptor tRNAs from bovine mitochondria by using a hybridization assay method.
For large scale preparation of mitochondrial tRNAs, a new hybridization assay method using synthetic oligodeoxyribonucleotide probes (16-17mer) complementary to individual tRNA sequences was developed and applied for the purification of two serine isoacceptor tRNAs (tRNASerAGY and tRNASerUCN) from bovine mitochondria. It is about 100 times more sensitive than the conventional aminoacylation assay method. 2-4 A260 units each of both tRNASer isoacceptors were purified from 17.5 kg of bovine liver, and they were characterized by means of nuclease digestion, melting profiles and aminoacylation activity. It is suggested that tRNASerUCN possesses the D loop/T loop interaction like usual L-shaped tRNAs, and that tRNASerAGY lacking almost an entire D arm is aminoacylated with an efficiency not very much lower than that of tRNASerUCN.
Yokogawa
T
T
Department of Industrial Chemistry, Faculty of Engineering, University of Tokyo, Japan.
Kumazawa
Y
Y
Miura
K
K
Watanabe
K
K
D000215:Acylation; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D008930:Mitochondria, Liver / Q000032:analysis*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012345:RNA, Transfer, Amino Acid-Specific / Q000302:isolation & purification*; D012362:RNA, Transfer, Ser / Q000302:isolation & purification* / Q000378:metabolism; D013329:Structure-Activity Relationship
D015345:Oligonucleotide Probes; D012345:RNA, Transfer, Amino Acid-Specific; D012362:RNA, Transfer, Ser