Abstract
Bacterial and eukaryotic transfer RNAs have been shown to contain hypermodified adenosine, 2-methylthio-N(6)-threonylcarbamoyladenosine, at position 37 (A(37)) adjacent to the 3'-end of the anticodon, which is essential for efficient and highly accurate protein translation by the ribosome. Using a combination of bioinformatic sequence analysis and in vivo assay coupled to HPLC/MS technique, we have identified, from distinct sequence signatures, two methylthiotransferase (MTTase) subfamilies, designated as MtaB in bacterial cells and e-MtaB in eukaryotic and archaeal cells. Both subfamilies are responsible for the transformation of N(6)-threonylcarbamoyladenosine into 2-methylthio-N(6)-threonylcarbamoyladenosine. Recently, a variant within the human CDKAL1 gene belonging to the e-MtaB subfamily was shown to predispose for type 2 diabetes. CDKAL1 is thus the first eukaryotic MTTase identified so far. Using purified preparations of Bacillus subtilis MtaB (YqeV), a CDKAL1 bacterial homolog, we demonstrate that YqeV/CDKAL1 enzymes, as the previously studied MTTases MiaB and RimO, contain two [4Fe-4S] clusters. This work lays the foundation for elucidating the function of CDKAL1.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine / analogs & derivatives*
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Adenosine / chemistry
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Adenosine / genetics
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Adenosine / metabolism
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Amino Acid Sequence
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Animals
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Archaea / enzymology
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Bacillus subtilis / enzymology
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Bacterial Proteins / chemistry*
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism
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Cyclin-Dependent Kinase 5 / chemistry*
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Cyclin-Dependent Kinase 5 / genetics
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Cyclin-Dependent Kinase 5 / metabolism
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Escherichia coli / enzymology
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Escherichia coli Proteins / chemistry*
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Escherichia coli Proteins / genetics
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Escherichia coli Proteins / metabolism
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Heat-Shock Proteins / chemistry*
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Heat-Shock Proteins / genetics
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Heat-Shock Proteins / metabolism
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Humans
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Mice
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Molecular Sequence Data
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RNA, Transfer / chemistry*
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RNA, Transfer / genetics
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RNA, Transfer / metabolism
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Sequence Analysis, Protein
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Sulfurtransferases / chemistry*
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Sulfurtransferases / genetics
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Sulfurtransferases / metabolism
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Threonine / analogs & derivatives*
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Threonine / chemistry
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Threonine / genetics
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Threonine / metabolism
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tRNA Methyltransferases
Substances
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2-methylthio-N6-threonylcarbamoyladenosine
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Bacterial Proteins
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Escherichia coli Proteins
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Heat-Shock Proteins
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yqeV protein, Bacillus subtilis
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Threonine
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RNA, Transfer
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tRNA Methyltransferases
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Cyclin-Dependent Kinase 5
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MiaB protein, E coli
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RimO protein, E coli
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Sulfurtransferases
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CDKAL1 protein, human
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Adenosine