title: Identification of Thermus thermophilus HB8 tRNA (Gm18) methyltransferase gene.
abstract: For the purpose of identification of the gene for Thermus thermophilus tRNA (Gm18) methyltransferase [tRNA (guanosine-2'-)-methyltransferase, EC 2.1.1.34], the purified enzyme from native source was analyzed by the peptide-mass mapping. The target gene encoded the amino acid sequences of the obtained peptides was searched in data from Thermus thermophilus HB8 genome-sequencing project. We found the target gene AB05130, which was expected to encode a protein composed of 194 amino acid residues and the molecular mass of this protein was calculated as 22083. The recombinant protein was expressed in E. coli as an active form. The Gm18 formation activity of the purified recombinant protein was confirmed by in vitro methylation followed by two-dimensional thin layer chromatography and Liquid Chromatography/Mass Spectrum analysis of substrate tRNA.
authors.affiliation: Dept. of Biochemistry and Mol. Biology, Ehime Univ.
authors.forename: H
authors.initials: H
authors.lastname: Hori
authors.forename: T
authors.initials: T
authors.lastname: Suzuki
authors.forename: K
authors.initials: K
authors.lastname: Sugawara
authors.forename: Y
authors.initials: Y
authors.lastname: Inoue
authors.forename: T
authors.initials: T
authors.lastname: Shibata
authors.forename: S
authors.initials: S
authors.lastname: Kuramitsu
authors.forename: S
authors.initials: S
authors.lastname: Yokoyama
authors.forename: T
authors.initials: T
authors.lastname: Oshima
authors.forename: K
authors.initials: K
authors.lastname: Watanabe
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D008745:Methylation; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D010449:Peptide Mapping; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012360:RNA, Transfer, Phe / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016963:Thermus thermophilus / Q000201:enzymology* / Q000235:genetics*; D012359:tRNA Methyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D012331:RNA, Fungal; D012360:RNA, Transfer, Phe; D011994:Recombinant Proteins; D012359:tRNA Methyltransferases; C023196:tRNA (guanine(10)-N(2))-dimethyltransferase