title: The dual identities of mammalian tRNA(Sec) for SerRS and selenocysteine synthase.
abstract: Se is an essential trace element and is found as a selenocysteine in the active site of Se-enzymes, such as glutathione peroxidase. tRNASec is first aminoacylated with serine by Ser RS and further is converted to selenocysteyl-tRNA by selenocysteine synthase. Mammalian selenocysteine tRNA has dual identities with Ser RS and selenocysteine synthase. Key identity elements for selenocysteine synthase are the long 9 bp AA- and long 6 bp D-stems. Major serine tRNA was converted to a mutant with a 9 bp AA-stem and 6 bp D-stem, instead of a 7 bp AA-stem and 3 bp D-stem. This mutant was active for selenylation as well as serylation. The relative kinetic parameter (Vmax/Km) of the mutant was 0.052 of the value (1.00) of wild-type Sec tRNA. This low value suggests that there is an unknown fine base specific for selenocysteine synthase. For serylation, mutant having 12 bp and wild type tRNASec having 13 bp of the total length of AA- + T-stems were active but the mutants having 11 or 14 bp were inactive. This shows that SerRS measures the distance between the discrimination base and long extra arm for recognition of tRNASer.
authors.affiliation: Faculty of Pharmaceutical Sciences, Nagoya City University, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Mizutani
authors.forename: K
authors.initials: K
authors.lastname: Kanaya
authors.forename: S
authors.initials: S
authors.lastname: Ikeda
authors.forename: T
authors.initials: T
authors.lastname: Fujiwara
authors.forename: K
authors.initials: K
authors.lastname: Yamada
authors.forename: T
authors.initials: T
authors.lastname: Totsuka
mesh_terms: D000818:Animals; D020029:Base Pairing; D001483:Base Sequence; D002417:Cattle; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008322:Mammals / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D017433:Protein Structure, Secondary; D012345:RNA, Transfer, Amino Acid-Specific / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012714:Serine-tRNA Ligase / Q000737:chemistry* / Q000378:metabolism; D014158:Transcription, Genetic; D014166:Transferases / Q000737:chemistry* / Q000378:metabolism
chemical_list: D012345:RNA, Transfer, Amino Acid-Specific; C036928:tRNA, selenocysteine-; D014166:Transferases; C060600:selenium transferase; D012714:Serine-tRNA Ligase