Preparation of biologically active Ascaris suum mitochondrial tRNAMet with a TV-replacement loop by ligation of chemically synthesized RNA fragments

Takashi Ohtsuki, Gota Kawai, Yoh Ichi Watanabe, Kiyoshi Kita, Kazuya Nishikawa, Kimitsuna Watanabe

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Ascaris suum mitochondrial tRNAMet lacking the entire T stem was prepared by enzymatic ligation of two chemically synthesized RNA fragments. The synthetic tRNA could be charged with methionine by A.suum mitochondrial extract, although the charging activity was considerably low compared with that of the native tRNA, probably due to lack of modification. Enzymatic probing of the synthetic tRNA showed a very similar digestion pattern to that of the native tRNAMet, which has already been concluded to take an L-shape-like structure [Watanabe et al. (1994) J. Biol. Chem., 269, 22902-22906]. These results suggest that the synthetic tRNA possesses almost the same conformation as the native one, irrespective of the presence or absence of modified residues. The method of preparing the bizarre tRNA used here will provide a useful tool for elucidating the tertiary structure of such tRNAs, because they can be obtained without too much difficulty in the amounts necessary for physicochemical studies such as NMR spectroscopy.

Original languageEnglish
Pages (from-to)662-667
Number of pages6
JournalNucleic acids research.
Volume24
Issue number4
Publication statusPublished - 1996
Externally publishedYes

Fingerprint

RNA, Transfer, Met
Ascaris suum
Transfer RNA
Ligation
RNA
Methionine
Digestion
Magnetic Resonance Spectroscopy

ASJC Scopus subject areas

  • Genetics

Cite this

Preparation of biologically active Ascaris suum mitochondrial tRNAMet with a TV-replacement loop by ligation of chemically synthesized RNA fragments. / Ohtsuki, Takashi; Kawai, Gota; Watanabe, Yoh Ichi; Kita, Kiyoshi; Nishikawa, Kazuya; Watanabe, Kimitsuna.

In: Nucleic acids research., Vol. 24, No. 4, 1996, p. 662-667.

Research output: Contribution to journalArticle

Ohtsuki, Takashi ; Kawai, Gota ; Watanabe, Yoh Ichi ; Kita, Kiyoshi ; Nishikawa, Kazuya ; Watanabe, Kimitsuna. / Preparation of biologically active Ascaris suum mitochondrial tRNAMet with a TV-replacement loop by ligation of chemically synthesized RNA fragments. In: Nucleic acids research. 1996 ; Vol. 24, No. 4. pp. 662-667.
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