GONUTS has been updated to MW1.31 Most things seem to be working but be sure to report problems.
Ny, T, Lindström, HR, Hagervall, TG and Björk, GR (1988) Purification of transfer RNA (m5U54)-methyltransferase from Escherichia coli. Association with RNA. Eur. J. Biochem. 177:467-75
tRNA (m5U54)-methyltransferase (EC 184.108.40.206) catalyzes the transfer of methyl groups from S-adenosyl-L-methionine to transfer ribonucleic acid (tRNA) and thereby forming 5-methyluridine (m5U, ribosylthymine) in position 54 of tRNA. This enzyme, which is involved in the biosynthesis of all tRNA chains in Escherichia coli, was purified 5800-fold. A hybrid plasmid carrying trmA, the structural gene for tRNA (m5U54)-methyltransferase was used to amplify genetically the production of this enzyme 40-fold. The purest fraction contained three polypeptides of 42 kDa, 41 kDa and 32 kDa and a heterogeneous 48-57-kDa RNA-protein complex. All the polypeptides seem to be related to the 42/41-kDa polypeptides previously identified as the tRNA (m5U54)-methyltransferase. RNA comprises about 50% (by mass) of the complex. The RNA seems not to be essential for the methylation activity, but may increase the activity of the enzyme. The amino acid composition is presented and the N-terminal sequence of the 42-kDa polypeptide was found to be: Met-Thr-Pro-Glu-His-Leu-Pro-Thr-Glu-Gln-Tyr-Glu-Ala-Gln-Leu-Ala-Glu-Lys- . The tRNA (m5U54)-methyltransferase has a pI of 4.7 and a pH optimum of 8.0. The enzyme does not require added cations but is stimulated by Mg2+. The apparent Km for tRNA and S-adenosyl-L-methionine are 80 nM and 17 microM, respectively.
Amino Acids/analysis; Chromatography, DEAE-Cellulose; Chromatography, Gel; Escherichia coli/enzymology; Escherichia coli/genetics; Gene Amplification; Genes; Genes, Bacterial; Kinetics; Plasmids; RNA, Bacterial/isolation & purification; tRNA Methyltransferases/genetics; tRNA Methyltransferases/isolation & purification; tRNA Methyltransferases/metabolism
|Gene product||Qualifier||GO Term||Evidence Code||with/from||Aspect||Extension||Notes||Status|
|GO:0008168: methyltransferase activity||
Figure 4 indicates the purified Gid protein catalyzes the site-specific formation of m5U-54 in [32P]-labeled yeast tRNAAsp transcript.
See Help:References for how to manage references in GONUTS.