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PMID: 28472312 Published · ppublish English Journal Article

ALKBH1 is an RNA dioxygenase responsible for cytoplasmic and mitochondrial tRNA modifications.

Nucleic acids research ·Vol. 45 ·No. 12 ·2017-07-07 ·Pages 7401-7415

Kawarada L, Suzuki T, Ohira T, Hirata S, Miyauchi K, Suzuki T

Abstract

ALKBH1 is a 2-oxoglutarate- and Fe2+-dependent dioxygenase responsible for multiple cellular functions. Here, we show that ALKBH1 is involved in biogenesis of 5-hydroxymethyl-2΄-O-methylcytidine (hm5Cm) and 5-formyl-2΄-O-methylcytidine (f5Cm) at the first position (position 34) of anticodon in cytoplasmic tRNALeu, as well as f5C at the same position in mitochondrial tRNAMet. Because f5C34 of mitochondrial tRNAMet is essential for translation of AUA, a non-universal codon in mammalian mitochondria, ALKBH1-knockout cells exhibited a strong reduction in mitochondrial translation and reduced respiratory complex activities, indicating that f5C34 formation mediated by ALKBH1 is required for efficient mitochondrial functions. We reconstituted formation of f5C34 on mitochondrial tRNAMetin vitro, and found that ALKBH1 first hydroxylated m5C34 to form hm5C34, and then oxidized hm5C34 to form f5C34. Moreover, we found that the frequency of 1-methyladenosine (m1A) in two mitochondrial tRNAs increased in ALKBH1-knockout cells, indicating that ALKBH1 also has demethylation activity toward m1A in mt-tRNAs. Based on these results, we conclude that nuclear and mitochondrial ALKBH1 play distinct roles in tRNA modification.

MeSH Terms
AlkB Homolog 1, Histone H2a Dioxygenase/deficiency,genetics Anticodon/chemistry,metabolism Base Sequence CRISPR-Cas Systems Cytidine/analogs & derivatives,metabolism Cytosol/metabolism Gene Knockout Techniques Genetic Complementation Test HEK293 Cells Humans Methyltransferases/genetics,metabolism Mitochondria/metabolism Nucleic Acid Conformation Oxidation-Reduction Oxidative Phosphorylation Protein Biosynthesis RNA, Transfer, Met/genetics,metabolism
Chemicals
2'-O-methyl-5-formylcytidine Anticodon RNA, Transfer, Met 5-formylcytidine Cytidine ALKBH1 protein, human AlkB Homolog 1, Histone H2a Dioxygenase Methyltransferases NSUN3 protein, human
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kawarada Layla
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Suzuki Takeo
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Ohira Takayuki
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Hirata Shoji
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Miyauchi Kenjyo
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Suzuki Tsutomu
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkx354
Published
2017-07-07
Pages
7401-7415
Language
English
Region
England
NLM ID
0411011
PMCID
PMC5499545
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/gkx354 PMC Full Text View journal details
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