State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
Institute of Urban Agriculture, Chinese Academy of Agricultural Sciences, Chengdu, China.
J Virol. 2022 Sep 14;96(17):e0077222. doi: 10.1128/jvi.00772-22. Epub 2022 Aug 16.
Bats are reservoirs for diverse coronaviruses, including swine acute diarrhea syndrome coronavirus (SADS-CoV). SADS-CoV was first identified in diarrheal piglets in 2017. As a novel alphacoronavirus, SADS-CoV shares ~95% identity with bat alphacoronavirus HKU2. SADS-CoV has been reported to have broad cell tropism and inherent potential to cross host species barriers for dissemination. Thus far, no effective antiviral drugs or vaccines are available to treat infections with SADS-CoV. Therefore, knowledge of the protein-coding gene set and a subcellular localization map of SADS-CoV proteins are fundamental first steps in this endeavor. Here, all SADS-CoV genes were cloned separately into Flag-tagged plasmids, and the subcellular localizations of viral proteins, with the exception of nsp11, were detected using confocal microscopy techniques. As a result, nsp1, nsp3-N, nsp4, nsp5, nsp7, nsp8, nsp9, nsp10, nsp14, and nsp15 were localized in the cytoplasm and nuclear spaces, and these viral proteins may perform specific functions in the nucleus. All structural and accessory proteins were mainly localized in the cytoplasm. NS7a and membrane protein M colocalized with the Golgi compartment, and they may regulate the assembly of SADS-CoV virions. Maturation of SADS-CoV may occur in the late endosomes, during which envelope protein E is involved in the assembly and release of the virus. In summary, the present study demonstrates for the first time the location of all the viral proteins of SADS-CoV. These fundamental studies of SADS-CoV will promote studies of basic virology of SADS-CoV and support preventive strategies for animals with infection of SADS-CoV. SADS-CoV is the first documented spillover of a bat coronavirus that causes severe diseases in domestic animals. Our study is an in-depth annotation of the newly discovered swine coronavirus SADS-CoV genome and viral protein expression. Systematic subcellular localization of SADS-CoV proteins can have dramatic significance in revealing viral protein biological functions in the subcellular locations. Furthermore, our study promote understanding the fundamental science behind the novel swine coronavirus to pave the way for treatments and cures.
蝙蝠是多种冠状病毒的宿主，包括猪急性腹泻综合征冠状病毒（SADS-CoV）。SADS-CoV 于 2017 年首次在腹泻仔猪中被发现。作为一种新型的甲型冠状病毒，SADS-CoV 与蝙蝠甲型冠状病毒 HKU2 有~95%的同源性。SADS-CoV 具有广泛的细胞嗜性和固有跨越宿主种间屏障传播的潜力。迄今为止，尚无有效的抗病毒药物或疫苗可用于治疗 SADS-CoV 感染。因此，了解 SADS-CoV 的蛋白编码基因集和病毒蛋白的亚细胞定位图谱是这一努力的基本第一步。在这里，我们将所有 SADS-CoV 基因分别克隆到 Flag 标记的质粒中，并使用共聚焦显微镜技术检测病毒蛋白的亚细胞定位，除 nsp11 外。结果表明，nsp1、nsp3-N、nsp4、nsp5、nsp7、nsp8、nsp9、nsp10、nsp14 和 nsp15 定位于细胞质和核空间，这些病毒蛋白可能在核内发挥特定功能。所有结构和辅助蛋白主要定位于细胞质中。NS7a 和膜蛋白 M 与高尔基体区室共定位，它们可能调节 SADS-CoV 病毒粒子的组装。SADS-CoV 的成熟可能发生在内体晚期，在此期间包膜蛋白 E 参与病毒的组装和释放。总之，本研究首次展示了 SADS-CoV 所有病毒蛋白的定位。这些 SADS-CoV 的基础研究将促进 SADS-CoV 基础病毒学的研究，并为感染 SADS-CoV 的动物提供预防策略。SADS-CoV 是第一种导致家畜严重疾病的蝙蝠冠状病毒的溢出。我们的研究是对新发现的猪冠状病毒 SADS-CoV 基因组和病毒蛋白表达的深入注释。SADS-CoV 蛋白的系统亚细胞定位在揭示病毒蛋白在亚细胞位置的生物学功能方面具有重要意义。此外，我们的研究促进了对新型猪冠状病毒的基础科学的理解，为治疗和治愈铺平了道路。
