PMID: 8604307Feb 15, 1996Paper

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

Nucleic Acids Research
T OhtsukiK Watanabe

Abstract

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.

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Citations

Oct 1, 2009·Journal of Biomolecular NMR·Kun LuMichael F Summers
Oct 20, 2005·Journal of Bioscience and Bioengineering·Katsuhiro YamanakaMasahiko Sisido
Jun 28, 2008·Nucleic Acids Research·Frank H T NelissenSybren S Wijmenga
Nov 7, 2003·Nucleic Acids Research·Shinya KurataKimitsuna Watanabe
Oct 6, 1998·Annual Review of Biochemistry·S Verma, F Eckstein
Jan 19, 2011·Progress in Nuclear Magnetic Resonance Spectroscopy·Cyril DominguezFrédéric H-T Allain
Feb 1, 2002·Nature Biotechnology·Ichiro HiraoShigeyuki Yokoyama
Mar 23, 2001·The Journal of Biological Chemistry·T OhtsukiK Watanabe
Feb 26, 2008·Molecular Biology and Evolution·Susan E Masta, Jeffrey L Boore

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