Nucleic Acids Research, Vol 24, Issue 4 662-667, Copyright © 1996 by Oxford University Press
T Ohtsuki, G Kawai, Y Watanabe, K Kita, K Nishikawa and K Watanabe
Ascaris suum mitochondrial tRNA Met 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 tRNA Met, 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.
ARTICLES
Preparation of biologically active Ascaris suum mitochondrial tRNAMet with a TV-replacement loop by ligation of chemically synthesized RNA fragments
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Japan.
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