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• W2072297370 abstract "Trypanosoma brucei is one of the most ancient eukaryotes where RNA interference (RNAi) is operational and is the only single-cell pathogen where RNAi has been extensively studied and used as a tool for functional analyses. Here, we report that the T. brucei RNAi pathway, although relying on a single Argonaute protein (AGO1), is initiated by the activities of two distinct Dicer-like enzymes. Both Tb DCL1, a mostly cytoplasmic protein, and the previously undescribed nuclear enzyme Tb DCL2 contribute to the biogenesis of siRNAs from retroposons. However, Tb DCL2 has a predominant role in generating siRNAs from chromosomal internal repeat transcripts that accumulate at the nucleolus in RNAi-deficient cells and in initiating the endogenous RNAi response against retroposons and repeats alike. Moreover, siRNAs generated by both Tb DCL1 and Tb DCL2 carry a 5′-monophosphate and a blocked 3′ terminus, suggesting that 3′ end modification is an ancient trait of siRNAs. We thus propose a model whereby Tb DCL2 fuels the T. brucei nuclear RNAi pathway and Tb DCL1 patrols the cytoplasm, posttranscriptionally silencing potentially harmful nucleic acid parasites that may access the cytoplasm. Nevertheless, we also provide evidence for cross-talk between the two Dicer-like enzymes, because Tb DCL2 is implicated in the generation of 35- to 65-nucleotide intermediate transcripts that appear to be substrates for Tb DCL1. Our finding that dcl2KO cells are more sensitive to RNAi triggers than wild-type cells has significant implications for reverse genetic analyses in this important human pathogen." @default.
• W2072297370 created "2016-06-24" @default.
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• W2072297370 date "2009-10-20" @default.
• W2072297370 modified "2023-09-23" @default.
• W2072297370 title "Distinct and overlapping roles for two Dicer-like proteins in the RNA interference pathways of the ancient eukaryote <i>Trypanosoma brucei</i>" @default.
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• W2072297370 doi "https://doi.org/10.1073/pnas.0907766106" @default.
• W2072297370 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2764927" @default.
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