Zhao Lijiu, Wei Xiangyu, Chen Fengyue, Chen Baoshan, Li Ru
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi Research Center for Microbial and Enzyme Engineering Technology, College of Life Science and Technology, Guangxi University, Nanning, China.
Guangxi Key Laboratory of Sugarcane Biology, College of Agriculture, Guangxi University, Nanning, China.
mBio. 2025 Jan 8;16(1):e0184424. doi: 10.1128/mbio.01844-24. Epub 2024 Nov 29.
As the most abundant eukaryotic mRNA modification, N-methyladenosine (mA) plays a crucial role in regulating multiple biological processes. This methylation is regulated by methyltransferases and demethylases. However, the regulatory role and mode of action of mA demethylases in fungi remain poorly understood. In this study, we demonstrate that CpALKBH is a demethylase in that removes mA modification from single-stranded RNA . The deletion of resulted in a significant increase in the mA methylation levels, along with decreases in the growth rate, sporulation, and virulence in . Additionally, CpZap1-a transcription factor-was identified as a downstream target of CpALKBH demethylase based on RNA sequencing analysis. We confirmed that CpALKBH demethylase regulates mRNA stability in an mA-dependent manner. Furthermore, through MazF assay, we found that methylation of at position 1935A is regulated by both CpALKBH demethylase and CpMTA1 methyltransferase. significantly influences the fungal phenotype and virulence, thereby restoring the abnormal phenotype observed in ∆ mutants. Collectively, our findings highlight the essential role of CpALKBH as an mA demethylase in the development and virulence of , while also elucidating the molecular mechanisms through which mA modification impacts mRNA stability.
N-methyladenosine (mA) is the most abundant eukaryotic mRNA modification and is involved in various biological processes. Methyltransferases and demethylases regulate the mA modification, but the regulatory role of mA demethylases in fungi remains poorly understood. Here, we demonstrated that CpALKBH functions as a demethylase in . The deletion of leads to a significant increase in mA levels and a reduction in fungal growth, sporulation, and virulence. We identified as a downstream target of CpALKBH, with CpALKBH regulating mRNA stability in an mA-dependent manner. Additionally, our findings indicate that methylation at position 1935A of is regulated by both the CpALKBH demethylase and the CpMTA1 methyltransferase. Given its critical role in fungal development and virulence, overexpression of can rescue abnormal phenotypes of ∆ mutant. Overall, these findings contribute to improving our understanding of the role of mA demethylase in fungi.
作为最丰富的真核生物mRNA修饰，N - 甲基腺苷（mA）在调节多种生物学过程中起关键作用。这种甲基化由甲基转移酶和去甲基酶调节。然而，mA去甲基酶在真菌中的调节作用和作用模式仍知之甚少。在本研究中，我们证明CpALKBH是一种去甲基酶，可从单链RNA上去除mA修饰。缺失该基因导致mA甲基化水平显著增加，同时生长速率、孢子形成和致病性降低。此外，基于RNA测序分析，CpZap1（一种转录因子）被鉴定为CpALKBH去甲基酶的下游靶点。我们证实CpALKBH去甲基酶以mA依赖的方式调节mRNA稳定性。此外，通过MazF分析，我们发现1935A位点的甲基化受CpALKBH去甲基酶和CpMTA1甲基转移酶共同调节。这显著影响真菌表型和致病性，从而恢复在∆突变体中观察到的异常表型。总体而言，我们的研究结果突出了CpALKBH作为mA去甲基酶在该真菌发育和致病性中的重要作用，同时也阐明了mA修饰影响mRNA稳定性的分子机制。
N - 甲基腺苷（mA）是最丰富的真核生物mRNA修饰，参与各种生物学过程。甲基转移酶和去甲基酶调节mA修饰，但mA去甲基酶在真菌中的调节作用仍知之甚少。在这里，我们证明CpALKBH在该真菌中起去甲基酶的作用。缺失该基因导致mA水平显著增加，真菌生长、孢子形成和致病性降低。我们鉴定出该基因是CpALKBH的下游靶点，CpALKBH以mA依赖的方式调节mRNA稳定性。此外，我们的研究结果表明，该基因1935A位点的甲基化受CpALKBH去甲基酶和CpMTA1甲基转移酶共同调节。鉴于其在真菌发育和致病性中的关键作用，该基因的过表达可挽救∆突变体的异常表型。总体而言，这些发现有助于增进我们对mA去甲基酶在真菌中作用的理解。
