Abstract
We have identified 12 initiator methionine tRNA (tRNAiMet) genes in the haploid human genome1. One gene, cloned from a recombinant phage library of human fetal liver DNA, contained a base substitution at position 56 in the mature eukaryotic initiator tRNA sequence. The substitution, a G replaced by a T, should generate a tRNA species having a U at a sequence position not observed in any of 178 sequenced prokaryotic and eukaryotic tRNAs2. Recently, we have demonstrated3 that this variant tRNA gene is efficiently transcribed in an homologous in vitro system derived from KB cells. However, the precursor RNA species of the variant gene, unlike that of the normal one, is not fully processed to a mature sequence in these cell-free extracts. Although the three nucleotides of the 5′ leader sequence are efficiently trimmed, the 13-nucleotide 3′ trailer is not processed, resulting in the accumulation of an 87-nucleotide precursor. In this report we assay the functional properties of the human variant initiator tRNA gene within an intact cell by studying its transcription from a simian virus 40 (SV40) recombinant during lytic infection of African green monkey kidney (AGMK) cells. We demonstrate that the variant gene is efficiently transcribed when inserted in the SV40 chromosome but, as in vitro, the precursor tRNA is not processed to a mature tRNA species.
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Zasloff, M., Santos, T. & Hamer, D. tRNA precursor transcribed from a mutant human gene inserted into a SV40 vector is processed incorrectly. Nature 295, 533–535 (1982). https://doi.org/10.1038/295533a0
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DOI: https://doi.org/10.1038/295533a0
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