Abstract
Trypanosomatidae is a family of early branching eukaryotes harbouring a distinctive repertoire of gene expression strategies. Functional mature messenger RNA is generated via the trans-splicing and polyadenylation processing of constitutively transcribed polycistronic units. Recently, trans-splicing of pre-small subunit ribosomal RNA in the 5' external transcribed spacer region and of precursor tRNAsec have been described. Here, we used a previously validated semi-nested reverse transcription-polymerase chain reaction strategy to investigate internal transcribed spacer (ITS) I acceptor sites in total RNA from Leishmania (Leishmania) amazonensis. Two distinct spliced leader-containing RNAs were detected indicating that trans-splicing reactions occur at two AG acceptor sites mapped in this ITS region. These data provide further evidence of the wide spectrum of RNA molecules that act as trans-splicing acceptors in trypanosomatids.
Highlights
Trypanosomatidae is a family of flagellated protozoans
spliced leader (SL) addition trans-splicing was exclusively associated with pre-messenger RNA (mRNA) processing and other RNA molecules were not considered to be involved in this processing mechanism
The nucleotide sequences at the 5’ ends of the two products were composed of the L. (L.) amazonensis SL forward primer followed by the downstream ME sequence linked to an internal transcribed spacer (ITS) I sequence
Summary
Trypanosomatidae is a family of flagellated protozoans. A variety of Trypanosomatidae species from the Leishmania and Trypanosoma genera are responsible for various human diseases that lead to millions of deaths in developing countries. We have identified pre-rRNA molecules that are processed by transsplicing in the 5’ external transcribed spacer (5’ETS) region in four trypanosomatid species [T. brucei, Trypanosoma cruzi, Leishmania (Leishmania) amazonensis and Crithidia fasciculate] and by 5’ETS polyadenylation in L.
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