The structure and repertoire of small interfering RNAs in Leishmania (Viannia) braziliensis reveal diversification in the trypanosomatid RNAi pathway

Summary Among trypanosomatid protozoa the mechanism of RNA interference (RNAi) has been investigated in Trypanosoma brucei and to a lesser extent in Leishmania braziliensis. Although these two parasitic organisms belong to the same family, they are evolutionarily distantly related raising questions...

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Veröffentlicht in:Molecular microbiology 2013-02, Vol.87 (3), p.580-593
Hauptverfasser: Atayde, Vanessa D., Shi, Huafang, Franklin, Joseph B., Carriero, Nicholas, Notton, Timothy, Lye, Lon‐Fye, Owens, Katherine, Beverley, Stephen M., Tschudi, Christian, Ullu, Elisabetta
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container_end_page 593
container_issue 3
container_start_page 580
container_title Molecular microbiology
container_volume 87
creator Atayde, Vanessa D.
Shi, Huafang
Franklin, Joseph B.
Carriero, Nicholas
Notton, Timothy
Lye, Lon‐Fye
Owens, Katherine
Beverley, Stephen M.
Tschudi, Christian
Ullu, Elisabetta
description Summary Among trypanosomatid protozoa the mechanism of RNA interference (RNAi) has been investigated in Trypanosoma brucei and to a lesser extent in Leishmania braziliensis. Although these two parasitic organisms belong to the same family, they are evolutionarily distantly related raising questions about the conservation of the RNAi pathway. Here we carried out an in‐depth analysis of small interfering RNAs (siRNAs) associated with L. braziliensis Argonaute1 (LbrAGO1). In contrast to T. brucei, Leishmania siRNAs are sensitive to 3′ end oxidation, indicating the absence of blocking groups, and the Leishmania genome does not code for a HEN1 RNA 2′‐O‐methyltransferase, which modifies small RNA 3′ ends. Consistent with this observation, ∼ 20% of siRNA 3′ ends carry non‐templated uridines. Thus siRNA biogenesis, and most likely their metabolism, is different in these organisms. Similarly to T. brucei, putative mobile elements and repeats constitute the major Leishmania siRNA‐producing loci and AGO1 ablation leads to accumulation of long transcripts derived from putative mobile elements. However, contrary to T. brucei, no siRNAs were detected from other genomic regions with the potential to form double‐stranded RNA, namely sites of convergent transcription and inverted repeats. Thus, our results indicate that organism‐specific diversification has occurred in the RNAi pathway during evolution of the trypanosomatid lineage.
doi_str_mv 10.1111/mmi.12117
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Although these two parasitic organisms belong to the same family, they are evolutionarily distantly related raising questions about the conservation of the RNAi pathway. Here we carried out an in‐depth analysis of small interfering RNAs (siRNAs) associated with L. braziliensis Argonaute1 (LbrAGO1). In contrast to T. brucei, Leishmania siRNAs are sensitive to 3′ end oxidation, indicating the absence of blocking groups, and the Leishmania genome does not code for a HEN1 RNA 2′‐O‐methyltransferase, which modifies small RNA 3′ ends. Consistent with this observation, ∼ 20% of siRNA 3′ ends carry non‐templated uridines. Thus siRNA biogenesis, and most likely their metabolism, is different in these organisms. Similarly to T. brucei, putative mobile elements and repeats constitute the major Leishmania siRNA‐producing loci and AGO1 ablation leads to accumulation of long transcripts derived from putative mobile elements. However, contrary to T. brucei, no siRNAs were detected from other genomic regions with the potential to form double‐stranded RNA, namely sites of convergent transcription and inverted repeats. 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subjects Argonaute Proteins - genetics
Bacteria
Biosynthesis
Gene Expression Regulation
Genetic Variation
Leishmania braziliensis
Leishmania braziliensis - genetics
Metabolism
Oxidation
Parasitic protozoa
Ribonucleic acid
RNA
RNA, Small Interfering - chemistry
RNA, Small Interfering - genetics
Trypanosoma brucei
Trypanosoma brucei brucei - genetics
Viannia braziliensis
title The structure and repertoire of small interfering RNAs in Leishmania (Viannia) braziliensis reveal diversification in the trypanosomatid RNAi pathway
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