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 language | English |
|---|---|
| Pages (from-to) | 662-667 |
| Number of pages | 6 |
| Journal | Nucleic acids research. |
| Volume | 24 |
| Issue number | 4 |
| Publication status | Published - 1996 |
| Externally published | Yes |
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- 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 journal › Article
}
TY - JOUR
T1 - Preparation of biologically active Ascaris suum mitochondrial tRNAMet with a TV-replacement loop by ligation of chemically synthesized RNA fragments
AU - Ohtsuki, Takashi
AU - Kawai, Gota
AU - Watanabe, Yoh Ichi
AU - Kita, Kiyoshi
AU - Nishikawa, Kazuya
AU - Watanabe, Kimitsuna
PY - 1996
Y1 - 1996
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0029991352&partnerID=8YFLogxK
M3 - Article
C2 - 8604307
AN - SCOPUS:0029991352
VL - 24
SP - 662
EP - 667
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 4
ER -