Thus, a major function of SjAgo2 appears to associate with the maintenance of genome stability via suppression of retroelements
Thus, a major function of SjAgo2 appears to associate with the maintenance of genome stability via suppression of retroelements. Table S3: Proteins identified by Orbitrap MS in different immunoprecipitates. (XLS) pntd.0001745.s007.xls (37K) GUID:?9E1BA673-EF74-4E65-A5C1-D824F94155D2 Table Pcdha10 S4: General information of the two small RNA libraries (SP1 and SP2). (XLS) pntd.0001745.s008.xls (20K) GUID:?E92F0309-A4F9-43E8-AF4F-9E008A90E311 Table S5: Data statistics of the two small RNA libraries (SP1 and SP2). (XLS) pntd.0001745.s009.xls (20K) GUID:?6F6D9B51-2F73-4656-BF56-C976FBA83B20 Abstract Background The complex life cycle of the genus Argonaute protein Ago2 (SjAgo2), but not SjAgo1 and SjAgo3, were generated. 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. The small RNA population associated with native SjAgo2 in adult parasites was extracted from the immunoprecipitated complex and Tipranavir subjected to library construction. High-through-put sequencing of these libraries yielded a total of 50 million high-quality reads. 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. 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. These results suggest that a key function of SjAgo2 is to maintain genome stability through suppressing the activities of retrotransposons. Conclusions/Significance In this study, we identified and characterized one of the three Argonautes, SjAgo2, and its associated small RNAs were found to be predominantly derived from particular classes of retrotransposons. Thus, a major function of SjAgo2 appears to associate with the maintenance of genome stability via suppression of retroelements. The data advance our understanding of the gene regulatory mechanisms in the blood fluke. Author Summary Schistosomiasis, a chronic disease caused by agents of the genus Argonaute proteins, SjAgo2, is involved in such mechanisms. 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. Tipranavir We found that SjAgo2 is mainly associated with particular types of retrotransposon-derived siRNAs. For instance, siRNAs generated from 10 classes of well-defined retrotransposons were significantly enriched in the SjAgo2-specific libraries. Thus, a major function of Ago2 in is proposed to be the maintenance of genome stability via retrotransposon suppression. Our findings advance understanding of the putative gene regulatory mechanisms in a flatworm parasite. Introduction Schistosomiasis is a chronic debilitating disease caused by the parasitic blood flukes of the genus to twenty-seven in the nematode (SjAgos) have been also reported by two groups [27], [28]. Both of them tried to determine the full-length sequences of the three Argonaute proteins and described the molecular characteristics of SjAgos. Chen during the parasite development and suggested that SjAgos coordinated in different SRRPs may be involved in regulating schistosome development [27]. In addition, no PIWI homologue was identified in were provided by Jiangxi Institute of Parasitic Diseases, Nanchang, China. The freshly released cercariae of were harvested for Total RNA isolation. To obtain hepatic schistosomula and adult worms, New Zealand White rabbits were percutaneously infected with cercariae (1000 to 1500 per rabbit). Hepatic schistosomula were isolated from the rabbits at 2 weeks post-infection, while mixed adult worms were obtained after 6-weeks post infection by hepatic-portal perfusion. Male and female adult worms were manually separated with the aid of a light microscope. Eggs were isolated from liver tissues of infected rabbits by enzyme digestion method [32]. All procedures performed on animals within Tipranavir this study were conducted following animal husbandry guidelines of the Chinese Academy Tipranavir of Medical Sciences and with permission from the Experimental Animal Committee of Chinese Academy of Medical Sciences with the Ethical Clearance Number IPB-2011-6. Total.