N⁶-methyladenosine (m⁶A) and N⁶,2'-O-dimethyladenosine (m⁶Am) modifications (m⁶A/m) of messenger RNA mediate diverse cellular functions. Oncogenic Kaposi's sarcoma-associated herpesvirus (KSHV) has latent and lytic replication phases that are essential for the development of KSHV-associated cancers. To date, the role of m⁶A/m in KSHV replication and tumorigenesis is unclear. Here, we provide mechanistic insights by examining the viral and cellular m⁶A/m epitranscriptomes during KSHV latent and lytic infection. KSHV transcripts contain abundant m⁶A/m modifications during latent and lytic replication, and these modifications are highly conserved among different cell types and infection systems. Knockdown of YTHDF2 enhanced lytic replication by impeding KSHV RNA degradation. YTHDF2 binds to viral transcripts and differentially mediates their stability. KSHV latent infection induces 5' untranslated region (UTR) hypomethylation and 3'UTR hypermethylation of the cellular epitranscriptome, regulating oncogenic and epithelial-mesenchymal transition pathways. KSHV lytic replication induces dynamic reprogramming of epitranscriptome, regulating pathways that control lytic replication. These results reveal a critical role of m⁶A/m modifications in KSHV lifecycle and provide rich resources for future investigations.

中文翻译：

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KSHV 生命周期中的病毒和细胞 N6-甲基腺苷和 N6,2'-O-二甲基腺苷外转录组。

信使RNA的N ⁶ -甲基腺苷(m ⁶ A)和N ⁶ ,2'-O-二甲基腺苷(m ⁶ Am)修饰(m ⁶ A/m)介导多种细胞功能。致癌卡波西肉瘤相关疱疹病毒 (KSHV) 具有潜伏期和裂解复制期，这对于 KSHV 相关癌症的发展至关重要。迄今为止，m ⁶ A/m 在 KSHV 复制和肿瘤发生中的作用尚不清楚。在这里，我们通过检查KSHV 潜伏和裂解感染期间的病毒和细胞 m ⁶ A/m 表观转录组来提供机制见解。KSHV 转录本含有丰富的 m ⁶潜伏和裂解复制过程中的 A/m 修饰，这些修饰在不同细胞类型和感染系统中高度保守。YTHDF2 的敲低通过阻止 KSHV RNA 降解增强了裂解复制。YTHDF2 与病毒转录本结合并以不同方式调节其稳定性。KSHV 潜伏感染诱导细胞表观转录组的 5' 非翻译区 (UTR) 低甲基化和 3'UTR 高甲基化，从而调节致癌和上皮间质转化途径。KSHV 裂解复制诱导表观转录组的动态重编程，调节控制裂解复制的途径。这些结果揭示了 m ⁶ A/m 修改在 KSHV 生命周期中的关键作用，并为未来的调查提供了丰富的资源。