![]() Our findings advance understanding of the putative gene regulatory mechanisms in a flatworm parasite.Ĭitation: Cai P, Piao X, Hou N, Liu S, Wang H, Chen Q (2012) Identification and Characterization of Argonaute Protein, Ago2 and Its Associated Small RNAs in Schistosoma japonicum. japonicum is proposed to be the maintenance of genome stability via retrotransposon suppression. For instance, siRNAs generated from 10 classes of well-defined retrotransposons were significantly enriched in the SjAgo2-specific libraries. We found that SjAgo2 is mainly associated with particular types of retrotransposon-derived siRNAs. By using specific mAb, native SjAgo2 protein was immunoisolated from a soluble adult worm antigen preparation, and its associated small RNAs were extracted for deep sequencing. japonicum Argonaute proteins, SjAgo2, is involved in such mechanisms. ![]() Our study showed that one of the three S. Thereby, the activity of these active mobile genetic elements should be restricted by elaborate mechanisms to protect genome stability. Transposable elements constitute a major component in the genome of schistosomes and have been recognized as remnants of evolutionary events, but some of them are still active today. The genomes of the three major pathogenic blood flukes, Schistosoma japonicum, Schistosoma mansoni, and Schistosoma haematobium, have been decoded as valuable entities for a systematic dissection of the biological characteristics of the parasites. Schistosomiasis, a chronic disease caused by agents of the genus Schistosoma, still afflicts more than 230 million people worldwide. These results suggest that a key function of SjAgo2 is to maintain genome stability through suppressing the activities of retrotransposons. Further bioinformatics analysis revealed that siRNAs derived from ten types of well-defined retrotransposons were dramatically enriched in the SjAgo2-specific libraries compared to small RNA libraries constructed with total small RNAs from separated adult worms. Classification of these small RNAs showed that endogenous siRNAs (endo-siRNAs) generated from transposable elements (TEs), especially from the subclasses of LINE and LTR, were prominent. High-through-put sequencing of these libraries yielded a total of ≈50 million high-quality reads. The small RNA population associated with native SjAgo2 in adult parasites was extracted from the immunoprecipitated complex and subjected to library construction. Soluble adult worm antigen preparation (SWAP) was subjected to immunoprecipitation with the mAbs and the captured SjAgo2 protein was subsequently confirmed by Western blot and mass spectrometry (MS) analysis. Two monoclonal antibodies (mAbs) specific to Schistosoma japonicum Argonaute protein Ago2 (SjAgo2), but not SjAgo1 and SjAgo3, were generated.
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