PMID:11779873
| Citation |
Agarwalla, S, Kealey, JT, Santi, DV and Stroud, RM (2002) Characterization of the 23 S ribosomal RNA m5U1939 methyltransferase from Escherichia coli. J. Biol. Chem. 277:8835-40 |
|---|---|
| Abstract |
An Escherichia coli open reading frame, ygcA, was identified as a putative 23 S ribosomal RNA 5-methyluridine methyltransferase (Gustafsson, C., Reid, R., Greene, P. J., and Santi, D. V. (1996) Nucleic Acids Res. 24, 3756-3762). We have cloned, expressed, and purified the 50-kDa protein encoded by ygcA. The purified enzyme catalyzed the AdoMet-dependent methylation of 23 S rRNA but did not act upon 16 S rRNA or tRNA. A high performance liquid chromatography-based nucleoside analysis identified the reaction product as 5-methyluridine. The enzyme specifically methylated U1939 as determined by a nuclease protection assay and by methylation assays using site-specific mutants of 23 S rRNA. A 40-nucleotide 23 S rRNA fragment (nucleotide 1930--1969) also served as an efficient substrate for the enzyme. The apparent K(m) values for the 40-mer RNA oligonucleotide and AdoMet were 3 and 26 microm, respectively, and the apparent k(cat) was 0.06 s(-1). The enzyme contains two equivalents of iron/monomer and has a sequence motif similar to a motif found in iron-sulfur proteins. We propose to name this gene rumA and accordingly name the protein product as RumA for RNA uridine methyltransferase. |
| Links |
PubMed Online version:10.1074/jbc.M111825200 |
| Keywords |
Bacterial Proteins/metabolism; Cloning, Molecular; Escherichia coli/enzymology; Methylation; RNA, Ribosomal, 23S/metabolism; Uridine/analogs & derivatives; Uridine/metabolism; tRNA Methyltransferases/genetics; tRNA Methyltransferases/isolation & purification; tRNA Methyltransferases/metabolism |
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