Abstract
We identified the first archaeal tRNA ribose 2'-O-methylase, aTrm56, belonging to the Cluster of Orthologous Groups (COG) 1303 that contains archaeal genes only. The corresponding protein exhibits a SPOUT S-adenosylmethionine (AdoMet)-dependent methyltransferase domain found in bacterial and yeast G18 tRNA 2'-O-methylases (SpoU, Trm3). We cloned the Pyrococcus abyssi PAB1040 gene belonging to this COG, expressed and purified the corresponding protein, and showed that in vitro, it specifically catalyzes the AdoMet-dependent 2'-O-ribose methylation of C at position 56 in tRNA transcripts. This tRNA methylation is present only in archaea, and the gene for this enzyme is present in all the archaeal genomes sequenced up to now, except in the crenarchaeon Pyrobaculum aerophilum. In this archaea, the C56 2'-O-methylation is provided by a C/D sRNP. Our work is the first demonstration that, within the same kingdom, two different mechanisms are used to modify the same nucleoside in tRNAs.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Catalysis
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Cloning, Molecular
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Consensus Sequence
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Cytosine / metabolism*
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Escherichia coli / genetics
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Genome, Archaeal
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Glutathione Transferase / metabolism
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Kinetics
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Molecular Sequence Data
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Molecular Weight
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Open Reading Frames
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Phylogeny
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Protein Structure, Secondary
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Pyrobaculum / genetics
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Pyrobaculum / metabolism
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Pyrococcus abyssi / enzymology
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Pyrococcus abyssi / genetics
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RNA, Archaeal / chemistry
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RNA, Archaeal / genetics
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RNA, Archaeal / metabolism*
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RNA, Small Nucleolar / genetics
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RNA, Small Nucleolar / metabolism*
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RNA, Transfer / chemistry*
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RNA, Transfer / metabolism
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Sequence Homology, Amino Acid
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Substrate Specificity
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Temperature
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tRNA Methyltransferases / chemistry*
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tRNA Methyltransferases / classification
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tRNA Methyltransferases / genetics
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tRNA Methyltransferases / metabolism*
Substances
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RNA, Archaeal
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RNA, Small Nucleolar
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Recombinant Proteins
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Cytosine
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RNA, Transfer
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tRNA Methyltransferases
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Glutathione Transferase