Division of Protective Immunity and Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, Philadelphia, PA, 19104, USA.
Department of Biomedical and Health Informatics, The Children's Hospital of Philadelphia, Philadelphia, PA, 19104, USA.
Nat Commun. 2020 Nov 26;11(1):6016. doi: 10.1038/s41467-020-19787-6.
Adenovirus is a nuclear replicating DNA virus reliant on host RNA processing machinery. Processing and metabolism of cellular RNAs can be regulated by METTL3, which catalyzes the addition of N6-methyladenosine (mA) to mRNAs. While mA-modified adenoviral RNAs have been previously detected, the location and function of this mark within the infectious cycle is unknown. Since the complex adenovirus transcriptome includes overlapping spliced units that would impede accurate mA mapping using short-read sequencing, here we profile mA within the adenovirus transcriptome using a combination of meRIP-seq and direct RNA long-read sequencing to yield both nucleotide and transcript-resolved mA detection. Although both early and late viral transcripts contain mA, depletion of mA writer METTL3 specifically impacts viral late transcripts by reducing their splicing efficiency. These data showcase a new technique for mA discovery within individual transcripts at nucleotide resolution, and highlight the role of mA in regulating splicing of a viral pathogen.
腺病毒是一种依赖于宿主 RNA 处理机制的核复制 DNA 病毒。METTL3 可以调节细胞 RNA 的加工和代谢，它催化 mRNAs 添加 N6-甲基腺苷（mA）。虽然以前已经检测到 mA 修饰的腺病毒 RNA，但在感染周期中这种修饰标记的位置和功能尚不清楚。由于复杂的腺病毒转录组包括重叠的拼接单元，这将阻碍使用短读测序进行准确的 mA 映射，因此我们使用 meRIP-seq 和直接 RNA 长读测序的组合来描绘腺病毒转录组中的 mA，以获得核苷酸和转录分辨率的 mA 检测。尽管早期和晚期病毒转录本都含有 mA，但 mA 书写器 METTL3 的耗竭特别通过降低其剪接效率来影响病毒晚期转录本。这些数据展示了一种在单个转录本中以核苷酸分辨率发现 mA 的新技术，并强调了 mA 在调节病毒病原体剪接中的作用。
