Isolation and characterization of the TRM1 locus, a gene essential for the N2,N2-dimethylguanosine modification of both mitochondrial and cytoplasmic tRNA in Saccharomyces cerevisiae.
The trm1 mutation of Saccharomyces cerevisiae is a single nuclear mutation that affects a specific base modification of both cytoplasmic and mitochondrial tRNA. Transfer RNA isolated from trm1 cells lacks the modified base N2,N2-dimethylguanosine, and extracts from these cells do not have detectable N2,N2-dimethylguanosine-specific tRNA methyltransferase activity. As part of our efforts to determine how this mutation affects enzyme activities in two different cellular compartments we have isolated the TRM1 locus by genetic complementation. The TRM1 locus restores the N2,N2-dimethylguanosine modification to both cytoplasmic and mitochondrial tRNA in trm1 cells. An open reading frame in this TRM1 gene is essential for complementation of the trm1 phenotype. Expression of this open reading frame in Escherichia coli converts the organism from one that neither makes N2,N2-dimethylguanosine nor has N2,N2-dimethylguanosine-specific tRNA methyltransferase activity into one that does. This result suggests that the TRM1 locus is the structural gene for the tRNA modification enzyme and that both nuclear/cytoplasmic and mitochondrial forms of the methyltransferase are produced from the same gene.
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D002874:Chromosome Mapping; D003593:Cytoplasm / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015247:Deoxyribonuclease HindIII; D015252:Deoxyribonucleases, Type II Site-Specific; D004926:Escherichia coli / Q000201:enzymology; D005796:Genes; D006151:Guanosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D008780:Methyltransferases / Q000378:metabolism; D009154:Mutation; D010641:Phenotype; D010957:Plasmids; D012313:RNA / Q000378:metabolism*; D000077278:RNA, Mitochondrial; D012343:RNA, Transfer / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D000077278:RNA, Mitochondrial; D006151:Guanosine; C008940:N(2),N(2)-dimethylguanosine; D012313:RNA; D012343:RNA, Transfer; D008780:Methyltransferases; D004262:DNA Restriction Enzymes; D015247:Deoxyribonuclease HindIII; C024406:endodeoxyribonuclease HpaI; D015252:Deoxyribonucleases, Type II Site-Specific