Analysis of genetic code ambiguity arising from nematode-specific misacylated tRNAs

Kiyofumi Hamashima, Masaru Mori, Yoshiki Andachi, Masaru Tomita, Yuji Kohara, Akio Kanai

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The faithful translation of the genetic code requires the highly accurate aminoacylation of transfer RNAs (tRNAs). However, it has been shown that nematode-specific V-arm-containing tRNAs (nev-tRNAs) are misacylated with leucine in vitro in a manner that transgresses the genetic code. nev-tRNA<sup>Gly</sup> (CCC) and nev-tRNA<sup>Ile</sup> (UAU), which are the major nev-tRNA isotypes, could theoretically decode the glycine (GGG) codon and isoleucine (AUA) codon as leucine, causing GGG and AUA codon ambiguity in nematode cells. To test this hypothesis, we investigated the functionality of nev-tRNAs and their impact on the proteome of Caenorhabditis elegans. Analysis of the nucleotide sequences in the 3' end regions of the nev-tRNAs showed that they had matured correctly, with the addition of CCA, which is a crucial posttranscriptional modification required for tRNA aminoacylation. The nuclear export of nev-tRNAs was confirmed with an analysis of their subcellular localization. These results show that nev-tRNAs are processed to their mature forms like common tRNAs and are available for translation. However, a whole-cell proteome analysis found no detectable level of nev-tRNA-induced mistranslation in C. elegans cells, suggesting that the genetic code is not ambiguous, at least under normal growth conditions. Our findings indicate that the translational fidelity of the nematode genetic code is strictly maintained, contrary to our expectations, although deviant tRNAs with misacylation properties are highly conserved in the nematode genome.

Original languageEnglish
Article numbere0116981
JournalPLoS One
Volume10
Issue number1
DOIs
Publication statusPublished - 2015 Jan 20

Fingerprint

Genetic Code
transfer RNA
genetic code
Transfer RNA
Nematoda
Transfer RNA Aminoacylation
Codon
codons
Caenorhabditis elegans
Proteome
Leucine
proteome
RNA, Transfer, Gly
translation (genetics)
leucine
RNA, Transfer, Ile
Cell Nucleus Active Transport
Isoleucine
cells
isoleucine

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Analysis of genetic code ambiguity arising from nematode-specific misacylated tRNAs. / Hamashima, Kiyofumi; Mori, Masaru; Andachi, Yoshiki; Tomita, Masaru; Kohara, Yuji; Kanai, Akio.

In: PLoS One, Vol. 10, No. 1, e0116981, 20.01.2015.

Research output: Contribution to journalArticle

Hamashima, Kiyofumi ; Mori, Masaru ; Andachi, Yoshiki ; Tomita, Masaru ; Kohara, Yuji ; Kanai, Akio. / Analysis of genetic code ambiguity arising from nematode-specific misacylated tRNAs. In: PLoS One. 2015 ; Vol. 10, No. 1.
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