ABSTRACT
Pre-mRNA maturation in trypanosomatids occurs through a process called trans-splicing which involves excision of introns and union of exons in two independent transcripts. For the first time, we present the standardization of Trypanosoma cruzi permeable cells (Y strain) as a model for trans-splicing study of mRNAs in trypanosomes, following by RNase protection reaction, which localizes the SL exon and intron. This trans-splicing reaction in vitro was also used to analyze the influence of NFOH-121, a nitrofurazone-derivative, on this mechanism. The results suggested that the prodrug affects the RNA processing in these parasites, but the trans-splicing reaction still occurred.
Subject(s)
Nitrofurazone/analogs & derivatives , Nitrofurazone/pharmacology , RNA Splicing/drug effects , RNA, Messenger/drug effects , RNA, Protozoan/drug effects , Trypanocidal Agents/pharmacology , Trypanosoma cruzi/genetics , Animals , Cell Membrane Permeability/drug effects , Exons/genetics , Introns/genetics , Time Factors , Transcription, Genetic/drug effects , Trypanosoma cruzi/drug effects , Trypanosoma cruzi/growth & developmentABSTRACT
Pre-mRNA maturation in trypanosomatids occurs through a process called trans-splicing which involves excision of introns and union of exons in two independent transcripts. For the first time, we present the standardization of Trypanosoma cruzi permeable cells (Y strain) as a model for trans-splicing study of mRNAs in trypanosomes, following by RNase protection reaction, which localizes the SL exon and intron. This trans-splicing reaction in vitro was also used to analyze the influence of NFOH-121, a nitrofurazone-derivative, on this mechanism. The results suggested that the prodrug affects the RNA processing in these parasites, but the trans-splicing reaction still occurred.
