Nucleic Acids Research, Vol 25, Issue 14 2694-2701, Copyright © 1997 by Oxford University Press
HQ Jiang, Y Motorin, YX Jin and H Grosjean
Cell-free yeast extract has been successfully used to catalyze the
enzymatic formation of 11 out of the 14 naturally occurring modified
nucleotides in yeast tRNAPhe(anticodon GAA). They are m2G10, D17, m22G26,
Cm32, Gm34,psi39, m5C40, m7G46, m5C49, T54 andpsi55. Only D16, Y37 and
m1A58 were not formed under in vitro conditions. However, m1G37was
quantitatively produced instead of Y37. The naturally occurring intron was
absolutely required for m5C40formation while it hindered completely the
enzymatic formation of Cm32, Gm34and m1G37. Enzymatic formation of
m22G26,psi39, m7G46, m5C49, T54 andpsi55were not or only slightly affected
by the presence of the intron. These results allow us to classify the
different tRNA modification enzymes into three groups: intron insensitive,
intron dependent, and those requiring the absence of the intron. The fact
that truncated tRNAPheconsisting of the anticodon stem and loop prolonged
with the 19 nucleotide long intron is a substrate for tRNA: cytosine-40
methylase demonstrates that the enzyme is not only strictly intron
dependent, but also does not require fully structured tRNA.
ARTICLES
Pleiotropic effects of intron removal on base modification pattern of yeast tRNAPhe: an in vitro study
Laboratoire d'Enzymologie et Biochimie Structurales, Centre National de la Recherche Scientifique, Avenue de la Terrasse, Batiment 34, F-91198 Gif-sur-Yvette, France.
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