Nucleic Acids Research, Vol 24, Issue 24 4987-4991, Copyright © 1996 by Oxford University Press
K Tomita, T Ueda and K Watanabe
In squid (Loligo bleekeri) mitochondria, the two 3'-terminal nucleotides
(G72-G73) of the tRNA(Tyr) gene overlap with the two 5'- terminal
nucleotides (G1-G2) of the downstream tRNA(Cys) gene. To elucidate the
processing mechanism(s) of the tRNA molecules derived from this region,
tRNAs were analyzed by sequencing cDNAs synthesized from circularized
tRNAs. Nucleotides G1-G2 in tRNA(Cys) appeared to be without
post-transcriptional conversion, whereas CCA was post- transcriptionally
added to the 3'-terminus. In contrast, in the majority of tRNAs(Tyr),
G72-G73 were found to be converted to A72-A73, accompanied by the CCA
addition. These results indicate that a precursor of tRNA(Tyr) is processed
at U71 and two adenosines are attached prior to the CCA addition. Thus, we
suggest that 5' processing of the precursor tRNA dominates 3' processing
and maturation of the tRNA is mediated by a polyadenylylation enzyme in the
mitochondria, a scenario which is consistent with the editing process
proposed in land snail mitochondria. We also obtained intermediates, such
as a premature tRNA lacking CCA that terminated at U71 and one with a
single adenosine attached at position 72, which support the suggested
maturation process. However, although we failed to detect a tRNA(Cys)
lacking G1- G2 at the 5'-terminus, we obtained cDNAs for tRNA(Tyr) with
G72-G73 and the CCA terminus. This inconsistent result suggests the
co-existence of another process(es) in the maturation of these tRNA
molecules in squid mitochondria.
ARTICLES
RNA editing in the acceptor stem of squid mitochondrial tRNA(Tyr)
Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo, Bunkyo-ku, Japan.
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