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| Status |
Public on Jul 19, 2021 |
| Title |
Depletion of m6A writers and readers impair 3'-end formation in Toxoplasma |
| Organism |
Toxoplasma gondii |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Toxoplasma gondii is an obligate intracellular parasite that can cause serious opportunistic disease in the immunocompromised or through congenital infection. To progress through its life cycle, Toxoplasma relies on multiple layers of gene regulation that includes an array of transcription and epigenetic factors. Over the last decade, the modification of mRNA has emerged as another important layer of gene regulation called epitranscriptomics. Here, we report that epitranscriptomics machinery exists in Toxoplasma, namely the methylation of adenosines (m6A) in mRNA transcripts. We identified novel components of the m6A methyltransferase complex and determined the distribution of m6A marks within the parasite transcriptome. m6A mapping revealed the modification to be preferentially located near the 3’-boundary of mRNAs within the consensus sequence, YGCAUGCR. Knockdown of the m6A writer components METTL3 and WTAP resulted in diminished m6A marks and a complete arrest of parasite replication. 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. Loss of METTL3, WTAP, or YTH1 lead to a defect in transcript 3’-end formation. Together, these findings establish that the m6A epitranscriptome is essential for parasite viability by contributing to the processing of mRNA 3’-ends.
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| Overall design |
expression changes in Toxoplasma upon IAA-induced knockdown of METTL3, WTAP, and YTH1 for 4 and 16 h
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| |
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| Contributor(s) |
Holmes MJ, Sullivan WJ Jr |
| Citation(s) |
34324585 |
| |
| Submission date |
Jun 16, 2021 |
| Last update date |
Aug 18, 2021 |
| Contact name |
Michael James Holmes |
| E-mail(s) |
[email protected]
|
| Organization name |
Indiana University School of Medicine
|
| Department |
Pharmacology & Toxicology
|
| Street address |
635 Barnhill Drive
|
| City |
Indianapolis |
| State/province |
IN |
| ZIP/Postal code |
46202 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL26742 |
Illumina NovaSeq 6000 (Toxoplasma gondii) |
|
| Samples (27)
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| Relations |
| BioProject |
PRJNA738537 |
| SRA |
SRP324453 |
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