title: Folate-/FAD-dependent tRNA methyltransferase from Thermus thermophilus regulates other modifications in tRNA at low temperatures.
abstract: TrmFO is a N(5) , N(10) -methylenetetrahydrofolate (CH2 THF)-/FAD-dependent tRNA methyltransferase, which synthesizes 5-methyluridine at position 54 (m(5) U54) in tRNA. Thermus thermophilus is an extreme-thermophilic eubacterium, which grows in a wide range of temperatures (50-83 °C). In T. thermophilus, modified nucleosides in tRNA and modification enzymes form a network, in which one modification regulates the degrees of other modifications and controls the flexibility of tRNA. To clarify the role of m(5) U54 and TrmFO in the network, we constructed the trmFO gene disruptant (∆trmFO) strain of T. thermophilus. Although this strain did not show any growth retardation at 70 °C, it showed a slow-growth phenotype at 50 °C. Nucleoside analysis showed increase in 2'-O-methylguanosine at position 18 and decrease in N(1) -methyladenosine at position 58 in the tRNA mixture from the ∆trmFO strain at 50 °C. These in vivo results were reproduced by in vitro experiments with purified enzymes. Thus, we concluded that the m(5) U54 modification have effects on the other modifications in tRNA through the network at 50 °C. (35) S incorporations into proteins showed that the protein synthesis activity of ∆trmFO strain was inferior to the wild-type strain at 50 °C, suggesting that the growth delay at 50 °C was caused by the inferior protein synthesis activity.
authors.affiliation: Department of Materials Science and Biotechnology, Graduate School of Science and Engineering, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
authors.forename: Ryota
authors.initials: R
authors.lastname: Yamagami
authors.affiliation: Department of Materials Science and Biotechnology, Graduate School of Science and Engineering, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
authors.forename: Chie
authors.initials: C
authors.lastname: Tomikawa
authors.affiliation: Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), 2-4-7 Aomi, Koto-ku, Tokyo, 135-0064, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Shigi
authors.affiliation: Department of Materials Science and Biotechnology, Graduate School of Science and Engineering, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
authors.forename: Ai
authors.initials: A
authors.lastname: Kazayama
authors.affiliation: Japan Biological Information Research Center (JBIRC), Japan Biological Informatics Consortium (JBIC), Tokyo, 135-0064, Japan.
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Asai
authors.affiliation: Department of Materials Science and Biotechnology, Graduate School of Science and Engineering, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Takuma
authors.affiliation: Department of Materials Science and Biotechnology, Graduate School of Science and Engineering, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Hirata
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), UMR9198, CEA, CNRS, Université Paris-Sud, 1 Avenue de la Terrasse, Gif-sur-Yvette, Cedex, 91198, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Fourmy
authors.affiliation: New England Biolabs, Inc, 240 County Road, Ipswich, MA, 01938, USA.
authors.forename: Haruichi
authors.initials: H
authors.lastname: Asahara
authors.affiliation: Biomedicinal Information Research Center (BIRC), National Institute of Advanced Industrial Science and Technology (AIST), 2-4-7 Aomi, Koto-ku, Tokyo, 135-0064, Japan.
authors.forename: Kimitsuna
authors.initials: K
authors.lastname: Watanabe
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), UMR9198, CEA, CNRS, Université Paris-Sud, 1 Avenue de la Terrasse, Gif-sur-Yvette, Cedex, 91198, France.
authors.forename: Satoko
authors.initials: S
authors.lastname: Yoshizawa
authors.affiliation: Department of Materials Science and Biotechnology, Graduate School of Science and Engineering, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Hori
mesh_terms: D005182:Flavin-Adenine Dinucleotide / Q000235:genetics / Q000378:metabolism; D005492:Folic Acid / Q000235:genetics / Q000378:metabolism; D006151:Guanosine / Q000031:analogs & derivatives / Q000235:genetics; D009154:Mutation; D012343:RNA, Transfer / Q000235:genetics*; D013696:Temperature; D016963:Thermus thermophilus / Q000201:enzymology / Q000235:genetics; D014529:Uridine / Q000031:analogs & derivatives / Q000235:genetics; D012359:tRNA Methyltransferases / Q000235:genetics* / Q000378:metabolism
chemical_list: D006151:Guanosine; D005182:Flavin-Adenine Dinucleotide; C009182:ribothymidine; D012343:RNA, Transfer; D005492:Folic Acid; D012359:tRNA Methyltransferases; C024900:2'-O-methylguanosine; D014529:Uridine