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- Title
N<sup>6</sup>-methyladenosine RNA methyltransferase CpMTA1 mediates CpAphA mRNA stability through a YTHDF1-dependent m<sup>6</sup>A modification in the chestnut blight fungus.
- Authors
Zhao, Lijiu; Wei, Xiangyu; Chen, Fengyue; Yuan, Luying; Chen, Baoshan; Li, Ru
- Abstract
In eukaryotic cells, N6-methyladenosine (m6A) is the most prevalent RNA epigenetic modification that plays crucial roles in multiple biological processes. Nevertheless, the functions and regulatory mechanisms of m6A in phytopathogenic fungi are poorly understood. Here, we showed that CpMTA1, an m6A methyltransferase in Cryphonectria parasitica, plays a crucial role in fungal phenotypic traits, virulence, and stress tolerance. Furthermore, the acid phosphatase gene CpAphA was implicated to be a target of CpMTA1 by integrated analysis of m6A-seq and RNA-seq, as in vivo RIP assay data confirmed that CpMTA1 directly interacts with CpAphA mRNA. Deletion of CpMTA1 drastically lowered the m6A level of CpAphA and reduced its mRNA expression. Moreover, we found that an m6A reader protein CpYTHDF1 recognizes CpAphA mRNA and increases its stability. Typically, the levels of CpAphA mRNA and protein exhibited a positive correlation with CpMTA1 and CpYTHDF1. Importantly, site-specific mutagenesis demonstrated that the m6A sites, A1306 and A1341, of CpAphA mRNA are important for fungal phenotypic traits and virulence in C. parasitica. Together, our findings demonstrate the essential role of the m6A methyltransferase CpMTA1 in C. parasitica, thereby advancing our understanding of fungal gene regulation through m6A modification. Author summary: Numerous modifications have been found in eukaryotic mRNA, with m6A appearing to be the predominant modification. However, the biological roles and regulatory mechanisms of these m6A-modified enzymes are still largely unknown in filamentous fungi. Here, we characterized an m6A methyltransferase, CPMTA1, in C. parasitica using a combined analysis of multiple phenotypes and multi-omics. Based on our results, CpMTA1 plays a crucial role in fungal phenotypic traits, virulence, and stress tolerance. Furthermore, m6A-seq and RNA-seq identified the acid phosphatase gene CpAphA as a target of CpMTA1.The m6A methylation of CpAphA was verified by MeRIP-qPCR and MazF analysis. We also found that an m6A reader protein CpYTHDF1 could recognize CpAphA mRNA and increase its stability. Importantly, the m6A sites A1306 and A1341 of CpAphA mRNA are involved in the regulation of fungal phenotypic traits and virulence in C. parasitica. These results provide a comprehensive insight into the function and mechanism of CpMTA1-mediated m6A modification in fungi, as well as improve our understanding of m6A-related studies.
- Subjects
RNA modification &; restriction; SITE-specific mutagenesis; GENE expression; ACID phosphatase; FUNGAL genes
- Publication
PLoS Pathogens, 2024, Vol 20, Issue 8, p1
- ISSN
1553-7366
- Publication type
Article
- DOI
10.1371/journal.ppat.1012476