Identification of eukaryotic and prokaryotic methylthiotransferase for biosynthesis of 2-methylthio-N6-threonylcarbamoyladenosine in tRNA.
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.
Arragain
Simon
S
Institut de Recherches en Technologie et Sciences pour le Vivant IRTSV-LCBM, UMR 5249 CEA/CNRS/UJF, Commissariat à l'Energie Atomique-Grenoble, 17 Avenue des Martyrs, 38054 Grenoble Cedex 09, France.
Handelman
Samuel K
SK
Forouhar
Farhad
F
Wei
Fan-Yan
FY
Tomizawa
Kazuhito
K
Hunt
John F
JF
Douki
Thierry
T
Fontecave
Marc
M
Mulliez
Etienne
E
Atta
Mohamed
M
D000241:Adenosine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001105:Archaea / Q000201:enzymology; D001412:Bacillus subtilis / Q000201:enzymology; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051360:Cyclin-Dependent Kinase 5 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D012343:RNA, Transfer / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020539:Sequence Analysis, Protein; D013466:Sulfurtransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013912:Threonine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012359:tRNA Methyltransferases
C515568:2-methylthio-N6-threonylcarbamoyladenosine; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; C104238:yqeV protein, Bacillus subtilis; D013912:Threonine; D012343:RNA, Transfer; D012359:tRNA Methyltransferases; D051360:Cyclin-Dependent Kinase 5; C499584:MiaB protein, E coli; C545011:RimO protein, E coli; D013466:Sulfurtransferases; C525110:CDKAL1 protein, human; D000241:Adenosine