African swine fever virus (ASFV) causes hemorrhagic fever in domestic pigs, presenting the biggest global threat to animal farming in recorded history. Despite the importance of ASFV, little is known about the mechanisms and regulation of ASFV transcription. Using RNA sequencing methods, we have determined total RNA abundance, transcription start sites, and transcription termination sites at single-nucleotide resolution. This allowed us to characterize DNA consensus motifs of early and late ASFV core promoters, as well as a polythymidylate sequence determinant for transcription termination. Our results demonstrate that ASFV utilizes alternative transcription start sites between early and late stages of infection and that ASFV RNA polymerase (RNAP) undergoes promoter-proximal transcript slippage at 5' ends of transcription units, adding quasitemplated AU- and AUAU-5' extensions to mRNAs. Here, we present the first much-needed genome-wide transcriptome study that provides unique insight into ASFV transcription and serves as a resource to aid future functional analyses of ASFV genes which are essential to combat this devastating disease.IMPORTANCE African swine fever virus (ASFV) causes incurable and often lethal hemorrhagic fever in domestic pigs. In 2020, ASF presents an acute and global animal health emergency that has the potential to devastate entire national economies as effective vaccines or antiviral drugs are not currently available (according to the Food and Agriculture Organization of the United Nations). With major outbreaks ongoing in Eastern Europe and Asia, urgent action is needed to advance our knowledge about the fundamental biology of ASFV, including the mechanisms and temporal control of gene expression. A thorough understanding of RNAP and transcription factor function, and of the sequence context of their promoter motifs, as well as accurate knowledge of which genes are expressed when and the amino acid sequence of the encoded proteins, is direly needed for the development of antiviral drugs and vaccines.

中文翻译：

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非洲猪瘟病毒转录组。

非洲猪瘟病毒 (ASFV) 引起家猪出血热，是有记录以来对畜牧业的最大全球威胁。尽管 ASFV 很重要，但对 ASFV 转录的机制和调控知之甚少。使用 RNA 测序方法，我们以单核苷酸分辨率确定了总 RNA 丰度、转录起始位点和转录终止位点。这使我们能够表征早期和晚期 ASFV 核心启动子的 DNA 共有基序，以及转录终止的聚胸苷酸序列决定因素。我们的研究结果表明，ASFV 在感染的早期和晚期利用替代转录起始位点，并且 ASFV RNA 聚合酶 (RNAP) 在转录单位的 5' 末端经历启动子近端转录物滑移，将准模板化的 AU- 和 AUAU-5' 延伸添加到 mRNA。在这里，我们提出了第一个急需的全基因组转录组研究，该研究提供了对 ASFV 转录的独特见解，并作为一种资源来帮助未来对 ASFV 基因进行功能分析，这些基因对于对抗这种毁灭性疾病至关重要。重要性非洲猪瘟病毒 (ASFV) ) 导致家猪出现无法治愈且通常致命的出血热。2020 年，非洲猪瘟呈现出严重的全球动物健康紧急情况，由于目前还没有有效的疫苗或抗病毒药物（根据联合国粮食及农业组织的数据），这有可能破坏整个国民经济。随着东欧和亚洲持续爆发大规模疫情，需要采取紧急行动来增进我们对 ASFV 基础生物学的了解，包括基因表达的机制和时间控制。全面了解 RNAP 和转录因子的功能，以及它们的启动子基序的序列背景，以及准确了解哪些基因何时表达以及所编码蛋白质的氨基酸序列，是开发抗病毒药物所迫切需要的和疫苗。