is also supported by a postdoctoral fellowship award from the American Heart Association (20POST35150019). This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All GEO files are available from the GEO database under accession numbers GSE165067 and GSE178355.įunding: This research was supported by grants from the National Institutes of Health (AI145239 and AI124723 to W.J.S.) and the Showalter Trust (to W.J.S.). Received: JanuAccepted: JPublished: July 29, 2021Ĭopyright: © 2021 Holmes et al. PLoS Pathog 17(7):Įditor: Dominique Soldati-Favre, University of Geneva, SWITZERLAND Together, these findings establish that the m6A epitranscriptome is essential for parasite viability by contributing to the processing of mRNA 3’-ends.Ĭitation: Holmes MJ, Padgett LR, Bastos MS, Sullivan WJ Jr (2021) m6A RNA methylation facilitates pre-mRNA 3’-end formation and is essential for viability of Toxoplasma gondii. Loss of METT元, WTAP, or YTH1 led to a defect in transcript 3’-end formation. Furthermore, we examined the two proteins in Toxoplasma that possess YTH domains, which bind m6A marks, and showed them to be integral members of the cleavage and polyadenylation machinery that catalyzes the 3’-end processing of pre-mRNAs. Knockdown of the m6A writer components METT元 and WTAP resulted in diminished m6A marks and a complete arrest of parasite replication. m6A mapping revealed the modification to be preferentially located near the 3’-boundary of mRNAs. We identified novel components of the m6A methyltransferase complex and determined the distribution of m6A marks within the parasite transcriptome. Here, we report that epitranscriptomics machinery exists in Toxoplasma, namely the methylation of adenosines (m6A) in mRNA transcripts. Over the last decade, the modification of mRNA has emerged as another important layer of gene regulation called epitranscriptomics.

To progress through its life cycle, Toxoplasma relies on multiple layers of gene regulation that includes an array of transcription and epigenetic factors. Toxoplasma gondii is an obligate intracellular parasite that can cause serious opportunistic disease in the immunocompromised or through congenital infection.

M6a dot blot protocol