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Comprehensive analysis of mRNA N6-methyladenosine modification in Eimeria tenella unsporulated oocysts and sporulated oocysts

ABSTRACT: N6-methyladenosine (m6A) modification, one of the most abundant RNA modification forms, is associated with almost all the aspects of RNA metabolism. In the present study, we surveyed the transcriptome-wide prevalence of m6A in sporulated oocysts and unsporulated oocysts of E. tenella. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) analysis showed that m6A modifications were most abundant in the coding sequence, followed by stop codon. There were 3903 hypermethylated and 3178 hypomethylated mRNAs in sporulated oocysts compared with unsporulated oocysts. Further joint analysis suggested that m6A modifications of the majority of genes were positively correlated with mRNA expression. The mRNA relative expression and m6A level of the selected genes were confirmed by quantitative reverse transcription PCR (RT-qPCR) and MeRIP-qPCR. GO and KEGG analysis indicated that differentially m6A methylated genes with significant differences in expression were closely related to processes such as regulation of gene expression, epigenetic, microtubule, autophagy-other and TOR signaling. Moreover, a total of 96 differentially expressed m6A genes without significant differences in expression showed significant differences at protein level. GO and pathway enrichment analysis of the 96 genes showed that RNA methylation may be involved in cell biosynthesis and metabolism of E. tenella.

ORGANISM(S): Eimeria tenella

PROVIDER: GSE221214 | GEO | 2022/12/22

REPOSITORIES: GEO

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Project description:Phosphorylation is a regulatory mechanism by which intracellular apicomplexan parasites control the process of invading and modifying host cells. However, the sporulated oocysts’ phosphoprotein repertoires between virulent and avirulent strains of Toxoplasma gondii and the underlying mechanisms contributing to virulence is still a mystery. A quantitative analysis of the sporulated oocysts’ phosphoproteome profile of T. gondii virulent and avirulent strains belonging to different genotypes was conducted to reveal phosphoprotein expression patterns that contribute to the virulence of a particular phenotype. Phosphopeptides from the sporulated oocysts of the virulent strain PYS (Chinese ToxoDB#9) and the avirulent strain type II (PRU strain) were enriched by titanium dioxide (TiO2) affinity chromatography and quantified using iBT technology. A total of 10,645 unique phosphopeptides, 8,181 nonredundant phosphorylation sites and 2,792 phosphoproteins were identified. The quantitative analysis identified 4,129 differentially expressed phosphopeptides (DEPs) between sporulated oocysts of the PYS strain and PRU strain (|log1.5 fold change| > 1 and P < 0.05), including 2,485 upregulated and 1,644 downregulated phosphopeptides. Motif analysis identified 24 motifs from the upregulated phosphorylated peptides including 22 serine motifs and two threonine motifs (TPE and TP), and 15 motifs from the downregulated phosphorylated peptides including 12 serine motifs and three threonine motifs (TP, RxxT and KxxT) in PYS strain when comparing PYS/PRU. Several kinases were consistent with motifs of overrepresented phosphopeptides, such as PKA, PKG, CKII, IKK, MAPK, EGFR, INSR, Jak, Syk, Src, Ab1. GO analysis, KEGG pathway analysis and STRING analysis revealed differentially expressed phosphoproteins (DEPs) exhibiting significant functional properties. Kinase associated network analysis showed that AGC kinase had the greatest connected peptides. Our findings reveal significant difference in phosphopeptides of sporulated oocysts between virulent and avirulent T. gondii strains, which provides solid foundation for further exploring the mechanisms contributing to the virulence of T. gondii.

Dataset Information

Comprehensive analysis of mRNA N6-methyladenosine modification in Eimeria tenella unsporulated oocysts and sporulated oocysts

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