title: Unexpected expansion of tRNA substrate recognition by the yeast m1G9 methyltransferase Trm10.
abstract: N-1 Methylation of the nearly invariant purine residue found at position 9 of tRNA is a nucleotide modification found in multiple tRNA species throughout Eukarya and Archaea. First discovered in Saccharomyces cerevisiae, the tRNA methyltransferase Trm10 is a highly conserved protein both necessary and sufficient to catalyze all known instances of m1G9 modification in yeast. Although there are 19 unique tRNA species that contain a G at position 9 in yeast, and whose fully modified sequence is known, only 9 of these tRNA species are modified with m1G9 in wild-type cells. The elements that allow Trm10 to distinguish between structurally similar tRNA species are not known, and sequences that are shared between all substrate or all nonsubstrate tRNAs have not been identified. Here, we demonstrate that the in vitro methylation activity of yeast Trm10 is not sufficient to explain the observed pattern of modification in vivo, as additional tRNA species are substrates for Trm10 m1G9 methyltransferase activity. Similarly, overexpression of Trm10 in yeast yields m1G9 containing tRNA species that are ordinarily unmodified in vivo. Thus, yeast Trm10 has a significantly broader tRNA substrate specificity than is suggested by the observed pattern of modification in wild-type yeast. These results may shed light onto the suggested involvement of Trm10 in other pathways in other organisms, particularly in higher eukaryotes that contain up to three different genes with sequence similarity to the single TRM10 gene in yeast, and where these other enzymes have been implicated in pathways beyond tRNA processing.
authors.affiliation: Center for RNA Biology, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: William E
authors.initials: WE
authors.lastname: Swinehart
authors.forename: Jeremy C
authors.initials: JC
authors.lastname: Henderson
authors.forename: Jane E
authors.initials: JE
authors.lastname: Jackman
mesh_terms: D005800:Genes, Fungal; D006151:Guanosine / Q000031:analogs & derivatives / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008745:Methylation; D012323:RNA Processing, Post-Transcriptional; D012331:RNA, Fungal / Q000737:chemistry / Q000378:metabolism*; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism*; D012366:RNA, Transfer, Val / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D012359:tRNA Methyltransferases / Q000235:genetics / Q000378:metabolism*
chemical_list: C116616:8-methylguanosine; D012331:RNA, Fungal; D012366:RNA, Transfer, Val; D029701:Saccharomyces cerevisiae Proteins; D006151:Guanosine; D012343:RNA, Transfer; C568603:TRM10 protein, S cerevisiae; D012359:tRNA Methyltransferases
keywords: Trm10; m1G; substrate specificity; tRNA methyltransferase