Key Laboratory of Animal Embryo Engineering and Molecular Breeding of Hubei Province, Institute of Animal Sciences and Veterinary Medicine, Hubei Academy of Agricultural Sciences, Wuhan 430070, China.
Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China.
Cells. 2024 Oct 21;13(20):1744. doi: 10.3390/cells13201744.
N6-methyladenosine (mA) is the most common modification in eukaryotic RNAs. Growing research indicates that mA methylation is crucial for a multitude of biological processes. However, research on the mA modifications in the regulation of porcine muscle growth is lacking. In this study, we identified differentially expressed genes in the neonatal period of muscle development between Large White (LW) and NingXiang (NX) pigs and further reported mA methylation patterns via MeRIP-seq. We found that mA modification regulates muscle cell development, myofibrils, cell cycle, and phosphatase regulator activity during the neonatal phase of muscle development. Interestingly, differentially expressed genes in LW and NX pigs were mainly enriched in pathways involved in protein synthesis. Furthermore, we performed a conjoint analysis of MeRIP-seq and RNA-seq data and identified 27 differentially expressed and mA-modified genes. Notably, a typical muscle-specific envelope transmembrane protein, WFS1, was differentially regulated by mA modifications in LW and NX pigs. We further revealed that the mA modification accelerated the degradation of WFS1 in a YTHDF2-dependent manner. Noteworthy, we identified a single nucleotide polymorphism (C21551T) within the last exon of WFS1 that resulted in variable mA methylation, contributing to the differing WFS1 expression levels observed in LW and NX pigs. Our study conducted a comprehensive analysis of the mA modification on NX and LW pigs during the neonatal period of muscle development, and elucidated the mechanism by which mA regulates the differential expression of WFS1 in the two breeds.
N6-甲基腺苷（mA）是真核 RNA 中最常见的修饰。越来越多的研究表明，mA 甲基化对于多种生物学过程至关重要。然而，关于 mA 修饰在猪肌肉生长调控中的研究还很缺乏。在这项研究中，我们鉴定了大白猪和宁乡猪肌肉发育新生儿期的差异表达基因，并通过 MeRIP-seq 进一步报道了 mA 甲基化模式。我们发现 mA 修饰调节肌肉细胞发育、肌原纤维、细胞周期和磷酸酶调节剂活性在肌肉发育的新生儿期。有趣的是，在 LW 和 NX 猪中差异表达的基因主要富集在与蛋白质合成相关的途径中。此外，我们对 MeRIP-seq 和 RNA-seq 数据进行了联合分析，鉴定了 27 个差异表达和 mA 修饰的基因。值得注意的是，一个典型的肌肉特异性包膜跨膜蛋白 WFS1，在 LW 和 NX 猪中受到 mA 修饰的差异调控。我们进一步揭示了 mA 修饰通过 YTHDF2 依赖性方式加速了 WFS1 的降解。值得注意的是，我们在 WFS1 的最后一个外显子中发现了一个单核苷酸多态性（C21551T），导致 mA 甲基化的可变，导致 LW 和 NX 猪中观察到的 WFS1 表达水平的不同。我们的研究对 NX 和 LW 猪在肌肉发育新生儿期的 mA 修饰进行了全面分析，并阐明了 mA 调节 WFS1 在两个品种中差异表达的机制。
