tRNA Synthesis & Processing
1. tRNA is transcribed by RNA polymerase III. The transcription product, the pre-tRNA, contains additional RNA sequences at both the 5’ and 3’-ends. These additional sequences are removed from the transcript during processing. The additional nucleotides at the 5’-end are removed by an unusual RNA containing enzyme called ribonuclease P (RNase P).
2. Some tRNA precursors contain an intron located in the anticodon arm. These introns are spliced out during processing of the tRNA.
3. All mature tRNAs contain the trinucleotide CCA at their 3’-end. These three bases are not coded for by the tRNA gene. Instead, these nucleotides are added during processing of the pre-tRNA transcript. The enzyme responsible for the addition of the CCA-end is tRNA nucleotidyl transferase and the reaction proceeds according to the following scheme:
tRNA +CTP --> tRNA-C + PPi
4. Mature tRNAs can contain up to 10% bases other than the usual adenine (A), guanine (G), cytidine (C) and uracil (U). These base modifications are introduced into the tRNA at the final processing step. The biological function of most of the modified bases is uncertain and the translation process seems normal in mutants lacking the enzymes responsible for modifying the bases.