Abstract
BackgroundProtein arginine methylation is a prevalent post-translational modification. The protein arginine methyltransferase family (PRMT) is involved in many cellular processes in eukaryotes, including transcriptional regulation, epigenetic regulation, RNA metabolism, and DNA damage repair. Toxoplasma gondii, an opportunistic protozoan parasite, encodes five conserved PRMTs. PRMT5 is thought to be responsible for substantial PRMT activity in T. gondii; however, it has not yet been characterized.MethodsWe tagged the 3′ end of the endogenous TgPRMT5 genomic locus with sequence encoding a 3X hemagglutinin (HA) epitope. IFA and WB were performed to check the expression and subcellular localization of TgPRMT5 in tachyzoites and bradyzoites. In vitro methylation assays were performed to determine whether endogenous TgPRMT5 has arginine methyltransferase activity.ResultsIFA and WB results showed that T. gondii PRMT5 (TgPRMT5) was localized in the cytoplasm in the tachyzoite stage; however, it shifts largely to the nuclear compartment in the bradyzoite stage. The in vitro methylation showed that TgPRMT5 has authentic type II PRMT activity and forms monomethylarginines and symmetric dimethylarginines.ConclusionsWe determined the expression and cellular localization of TgPRMT5 in tachyzoites and bradyzoites and confirmed its type II PRMT activity. We demonstrated the major changes in expression and cellular localization of TgPRMT5 during the tachyzoite and bradyzoite stages in T. gondii. Our findings suggest that TgPRMT5 protein may be involved in tachyzoite-bradyzoite transformation.
Highlights
Protein arginine methylation is a prevalent post-translational modification
Subcellular localization of T. gondii PRMT5 (TgPRMT5) immunofluorescent assay (IFA) was performed to check subcellular localization of TgPRMT5, and the TgBAG1 was expressed in the cytoplasm in the bradyzoite stage as a bradyzoite marker
IFA results showed that TgPRMT5 was localized in the cytoplasm in the tachyzoite stage; it was mainly localized in the nucleus in the bradyzoite stage (Fig. 2a)
Summary
Protein arginine methylation is a prevalent post-translational modification. The protein arginine meth‐ yltransferase family (PRMT) is involved in many cellular processes in eukaryotes, including transcriptional regulation, epigenetic regulation, RNA metabolism, and DNA damage repair. Toxoplasma gondii (T. gondii) is an obligate intracellular parasite of the phylum Apicomplexa. This species is the causative agent of toxoplasmosis, which is estimated to infect 16–40% of the population of the USA and up to 80% of the population in other countries [1]. Stage-specific gene expression is controlled by the concerted actions of specific transcription factors and repressors, translational repression, and epigenetic mechanisms [2]. Significant alterations in the expressed transcriptome and remodeling of chromatin structure, a major mechanism restricting and regulating access to genomic DNA, are occurring during the conversion between the tachyzoite and bradyzoite. Characterization of the T. gondii histone-modifying enzymes will provide a better understanding of the role of epigenetic regulation in gene expression in pathogenic apicomplexan parasites like T. gondii
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