Effects of Trypanosoma brucei tryptophanyl-tRNA synthetases silencing by RNA interference

The kinetoplast genetic code deviates from the universal code in that 90% of mitochondrial tryptophans are specified by UGA instead of UGG codons. A single nucleus-encoded tRNA Trp(CCA) is used by both nuclear and mitochondria genes, since all kinetoplast tRNAs are imported into the mitochondria from the cytoplasm. To allow decoding of the mitochondrial UGA codons as tryptophan, the tRNA Trp(CCA) anticodon is changed to UCA by an editing event. Two tryptophanyl tRNA synthetases (TrpRSs) have been identified in Trypanosoma brucei: TbTrpRS1 and TbTrpRS2 which localize to the cytoplasm and mitochondria respectively. We used inducible RNA interference (RNAi) to assess the role of TbTrpRSs. Our data validates previous observations of TrpRS as potential drug design targets and investigates the RNAi effect on the mitochondria of the parasite.     

RNA polymerase I-mediated protein-coding gene expression in Trypanosoma brucei

Protein-coding genes are transcribed by RNA polymerise (pol) II in all eukaryotes analyzed to date, with the exception of the protozoan Trypanosoma brucei, where pol I can mediate expression of chloramphenicol acetyl transferase (CAT) and neomycin phosphotransferase (neo) reporter genes. The addition of the capped 39-nucleotide (nt) mini-exon to the pre-messenger RNA (mRNA) by trans-splicing in T. brucei has presumably led to the uncoupling of the requirement for production of mRNA by pol II. Here Hui-min Chung, Mary G-S. Lee and Lex Van der Ploeg review the evidence that supports the notion that pol I also transcribes a subset of naturally occurring protein-coding genes in T. brucei.     

Complete sequence of the gene encoding the largest subunit of RNA polymerase I of Trypanosoma brucei

We have set out to clone the trypanosomal gene encoding the largest subunit of RNA polymerase I. We screened a genomic library with a synthetic oligonucleotide probe encoding an eleven amino acid sequence motif, YNADFDGDEMN, which has been found in all eukaryotic RNA polymerase largest subunit genes analyzed so far. We isolated the Trp11 locus and determined the complete sequence of the gene encoded within this locus. The deduced amino acid sequence contains the highly conserved RNA polymerase domains as well as the previously identified RNA polymerase I-specific hydrophilic insertions. Therefore, the gene most closely resembles the largest subunit of RNA polymerase I.