Hostname: page-component-586b7cd67f-r5fsc Total loading time: 0 Render date: 2024-11-23T01:49:16.879Z Has data issue: false hasContentIssue false

The yeast Saccharomyces cerevisiae YDL112w ORF encodes the putative 2′-O-ribose methyltransferase catalyzing the formation of Gm18 in tRNAs

Published online by Cambridge University Press:  01 January 1999

JÉRÔME CAVAILLÉ
Affiliation:
Laboratoire de Biologie Moléculaire Eucaryote du C.N.R.S., Université Paul-Sabatier, 118 route de Narbonne, 31062 Toulouse Cédex, France
FARID CHETOUANI
Affiliation:
Laboratoire de Biologie Moléculaire Eucaryote du C.N.R.S., Université Paul-Sabatier, 118 route de Narbonne, 31062 Toulouse Cédex, France
JEAN-PIERRE BACHELLERIE
Affiliation:
Laboratoire de Biologie Moléculaire Eucaryote du C.N.R.S., Université Paul-Sabatier, 118 route de Narbonne, 31062 Toulouse Cédex, France
Get access

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.

Type
Research Article
Copyright
© 1999 RNA Society

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)