tRNA His maturation: An essential yeast protein catalyzes addition of a guanine nucleotide to the 5 (cid:1) end of tRNA His

All tRNA His molecules are unusual in having an extra 5 (cid:1) GMP residue (G −1 ) that, in eukaryotes, is added after transcription and RNase P cleavage. Incorporation of this G −1 residue is a rare example of nucleotide addition occurring at an RNA 5 (cid:1) end in a normal phosphodiester linkage. We show here that the essential Saccharomyces cerevisiae ORF YGR024c ( THG1 ) is responsible for this guanylyltransferase reaction. Thg1p was identified by survey of a genomic collection of yeast GST-ORF fusion proteins for addition of [ (cid:1) - 32 P]GTP to tRNA His . End analysis confirms the presence of G −1 . Thg1p is required for tRNA His guanylylation in vivo, because cells depleted of Thg1p lack G −1 in their tRNA His . His 6 -Thg1p purified from Escherichia coli catalyzes the guanylyltransferase step of G −1 addition using a ppp-tRNA His substrate, and appears to catalyze the activation step using p-tRNA His and ATP. Thg1p is highly conserved in eukaryotes, where G −1 addition is necessary, and is not found in eubacteria, where G −1 is genome-encoded. Thus, Thg1p is the first member of a new family of enzymes that can catalyze phosphodiester bond formation at the 5 (cid:1) end of RNAs, formally in a 3 (cid:1) –5 (cid:1) direction. Surprisingly, despite its varied activities, Thg1p contains no recognizable catalytic or functional domains.

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