Hetero subunit interaction and RNA recognition of yeast tRNA (m7G46) methyltransferase synthesized in a wheat germ cell-free translation system.
Yeast tRNA (m(7)G46) methyltransferase contains two protein subunits (Trm8 and Trm82). The enzyme catalyzes a methyl-transfer from S-adenosyl-L-methionine to the N(7) atom of guanine at position 46 in tRNA. We deviced synthesis of active Trm8-Trm82 heterodimer in a wheat germ cell-free translation system. When Trm8 or Trm82 mRNA were used for a synthesis, Trm8 or Trm82 protein could be synthesized. Upon mixing the synthesized Trm8 and Trm82 proteins, no active Trm8-Trm82 heterodimer was produced. Active Trm8-Trm82 heterodimer was only synthesized under conditions, in which both Trm8 and Trm82 mRNAs were co-translated. To address the RNA recognition mechanism of the Trm8-Trm82 complex, we investigated methyl acceptance activities of eight truncated yeast tRNA(Phe) transcripts. In this meeting, we demonstrate that yeast Trm8-Trm82 has stricter recognition requirements for the tRNA molecule as compared to the bacterial enzyme, TrmB.
Muneyoshi
Yuki
Y
Department of Materials and Biotechnology, Graduate School of Science and Engineering Ehime University, Matsuyama, 790-8577 Japan.
Matsumoto
Keisuke
K
Tomikawa
Chie
C
Toyooka
Takashi
T
Ochi
Anna
A
Masaoka
Takashi
T
Endo
Yaeta
Y
Hori
Hiroyuki
H
D001419:Bacteria / Q000201:enzymology; D001483:Base Sequence; D002474:Cell-Free System; D019281:Dimerization; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D021122:Protein Subunits / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012360:RNA, Transfer, Phe / Q000737:chemistry / Q000378:metabolism; D012639:Seeds / Q000235:genetics; D014908:Triticum / Q000196:embryology / Q000235:genetics; D015003:Yeasts / Q000201:enzymology*; D012359:tRNA Methyltransferases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
D005656:Fungal Proteins; D021122:Protein Subunits; D012360:RNA, Transfer, Phe; D012359:tRNA Methyltransferases; C036814:tRNA (guanine-N7-)-methyltransferase