Cdk5rap1-mediated 2-methylthio-N6-isopentenyladenosine modification is absent from nuclear-derived RNA species

Md Fakruddin, Fan Yan Wei, Shohei Emura, Shigeru Matsuda, Takehiro Yasukawa, Dongchon Kang, Kazuhito Tomizawa

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

2-Methylthio-N6-isopentenyl modification of adenosine (ms2i6A) is an evolutionally conserved modification that is found in transfer RNAs (tRNAs). We have recently shown that Cdk5 regulatory subunit-associated protein 1 (Cdk5rap1) specifically converts i6A to ms2i6A at position A37 of four mitochondrial DNA-encoded tRNAs, and that the modification regulates efficient mitochondrial translation and energy metabolism in mammals. Curiously, a previous study reported that ms2i6A is present abundantly in nuclear-derived RNA species such as microRNAs, but not in tRNA fractions. To fully understand the molecular property of ms2i6A, the existence of non-canonical ms2i6A must be carefully validated. In the present study, we examined ms2i6A in total RNA purified from human and murine ρ0 cells, in which mitochondrial DNA-derived tRNAs were completely depleted. The ms2i6A was not detected in these cells at all. We generated a monoclonal antibody against ms2i6A and examined ms2i6A in murine RNAs using the antibody. The anti-ms2i6A antibody only reacted with the tRNA fractions and not in other RNA species. Furthermore, immunocytochemistry analysis using the antibody showed the predominant localization of ms2i6A in mitochondria and co-localization with the mitochondrial elongation factor Tu. Taken together, we propose that ms2i6A is a mitochondrial tRNA-specific modification and is absent from nuclear-encoded RNA species.

Original languageEnglish
Pages (from-to)11954-11961
Number of pages8
JournalNucleic Acids Research
Volume45
Issue number20
DOIs
Publication statusPublished - Nov 16 2017
Externally publishedYes

Fingerprint

Isopentenyladenosine
Nuclear RNA
Protein Subunits
Transfer RNA
RNA
Mitochondrial DNA
Peptide Elongation Factors
Antibodies
MicroRNAs
Adenosine
Energy Metabolism
Mammals
Anti-Idiotypic Antibodies
Mitochondria
Immunohistochemistry
Monoclonal Antibodies

ASJC Scopus subject areas

  • Genetics

Cite this

Fakruddin, M., Wei, F. Y., Emura, S., Matsuda, S., Yasukawa, T., Kang, D., & Tomizawa, K. (2017). Cdk5rap1-mediated 2-methylthio-N6-isopentenyladenosine modification is absent from nuclear-derived RNA species. Nucleic Acids Research, 45(20), 11954-11961. https://doi.org/10.1093/nar/gkx819

Cdk5rap1-mediated 2-methylthio-N6-isopentenyladenosine modification is absent from nuclear-derived RNA species. / Fakruddin, Md; Wei, Fan Yan; Emura, Shohei; Matsuda, Shigeru; Yasukawa, Takehiro; Kang, Dongchon; Tomizawa, Kazuhito.

In: Nucleic Acids Research, Vol. 45, No. 20, 16.11.2017, p. 11954-11961.

Research output: Contribution to journalArticle

Fakruddin, M, Wei, FY, Emura, S, Matsuda, S, Yasukawa, T, Kang, D & Tomizawa, K 2017, 'Cdk5rap1-mediated 2-methylthio-N6-isopentenyladenosine modification is absent from nuclear-derived RNA species', Nucleic Acids Research, vol. 45, no. 20, pp. 11954-11961. https://doi.org/10.1093/nar/gkx819
Fakruddin, Md ; Wei, Fan Yan ; Emura, Shohei ; Matsuda, Shigeru ; Yasukawa, Takehiro ; Kang, Dongchon ; Tomizawa, Kazuhito. / Cdk5rap1-mediated 2-methylthio-N6-isopentenyladenosine modification is absent from nuclear-derived RNA species. In: Nucleic Acids Research. 2017 ; Vol. 45, No. 20. pp. 11954-11961.
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