The yeast Saccharomyces cerevisiae YDL112w ORF encodes the putative 2′-O-ribose methyltransferase catalyzing the formation of Gm18 in tRNAs
- 1 January 1999
- journal article
- research article
- Published by Cold Spring Harbor Laboratory in RNA
- Vol. 5 (1), 66-81
- https://doi.org/10.1017/s1355838299981475
Abstract
The protein sequences of three known RNA 2′-O-ribose methylases were used as probes for detecting putative homologs through iterative searches of genomic databases. We have identified 45 new positive Open Reading Frames (ORFs), mostly in prokaryotic genomes. Five complete eukaryotic ORFs were also detected, among which was a single ORF (YDL112w) in the yeast Saccharomyces cerevisiae genome. After genetic depletion of YDL112w, we observed a specific defect in tRNA ribose methylation, with the complete disappearance of Gm18 in all tRNAs that naturally contain this modification, whereas other tRNA ribose methylations and the complex pattern of rRNA ribose methylations were not affected. The tRNA G18 methylation defect was suppressed by transformation of the disrupted strain with a plasmid allowing expression of YDL112wp. The formation of Gm18 on an in vitro transcript of a yeast tRNASer naturally containing this methylation, which was efficiently catalyzed by cell-free extracts from the wild-type yeast strain, did not occur with extracts from the disrupted strain. The protein encoded by the YDL112w ORF, termed Trm3 (tRNA methylation), is therefore likely to be the tRNA (Gm18) ribose methylase. In in vitro assays, its activity is strongly dependent on tRNA architecture. Trm3p, the first putative tRNA ribose methylase identified in an eukaryotic organism, is considerably larger than its Escherichia coli functional homolog spoU (1,436 amino acids vs. 229 amino acids), or any known or putative prokaryotic RNA ribose methyltransferase. Homologs found in human (TRP-185 protein), Caenorhabditis elegans and Arabidopsis thaliana also exhibit a very long N-terminal extension not related to any protein sequence in databases.This publication has 72 references indexed in Scilit:
- Methylation of mRNAPublished by Wiley ,2006
- Separate information required for nuclear and subnuclear localization: additional complexity in localizing an enzyme shared by mitochondria and nuclei.Molecular and Cellular Biology, 1992
- Methylation sites in Escherichia coli ribosomal RNA: Localization and identification of four new sites of methylation in 23 S rRNABiochemistry, 1992
- The gene for a tRNA modifying enzyme, m5U54-methyltransferase, is essential for viability in Escherichia coli.Proceedings of the National Academy of Sciences of the United States of America, 1992
- Two distinct nuclear transcription factors recognize loop and bulge residues of the HIV-1 TAR RNA hairpin.Genes & Development, 1991
- tat regulates binding of the human immunodeficiency virus trans-activating region RNA loop-binding protein TRP-185.Genes & Development, 1991
- A family of low and high copy replicative, integrative and single‐stranded S. cerevisiae/E. coli shuttle vectorsYeast, 1991
- [30] Labeling of RNA and phosphoproteins in Saccharomyces cerevisiaeMethods in Enzymology, 1991
- Aminoacyl-tRNA Synthetase Family from Prokaryotes and Eukaryotes: Structural Domains and Their ImplicationsProgress in Nucleic Acid Research and Molecular Biology, 1991
- 2'-O-Methylated oligonucleotides in ribosomal 18S and 28S RNA of a mouse hepatoma, MH 134Biochemistry, 1975