Banyangvirus is a new genus (Phenuiviridae family, Bunyavirales order) that comprises a group of emerging tick-borne viruses with severe fever with thrombocytopenia syndrome virus (SFTSV) and Heartland virus (HRTV) as virulent representatives. As segmented RNA viruses, bunyaviruses may have genome reassortment potential, increasing the concern about new life-threatening bunyavirus emergence. Using a series of combinatory minigenome reporter assays based on transfection and superinfection, we showed that replication machinery proteins of designated banyangviruses can recognize genomic untranslated regions (UTRs) of other banyangviruses and assemble heterogenous minigenomes into functional ribonucleoproteins (RNPs). Moreover, both heterogenous and heterozygous RNPs were efficiently packaged by viral glycoproteins into infectious virus-like particles, manifesting remarkable reassortment potential of banyangviruses. Meanwhile, UTR promoter strength of the three banyangvirus segments appeared to be M > L > S. Secondary structure analysis revealed a conservative non-basepairing protruding nucleotide in the terminal UTR panhandles of M and L (but not S) segments of all banyangviruses and some related phleboviruses (Phlebovirus genus). Furthermore, not only a conserved panhandle region but also the protruding nucleotide proved important for UTR function. Removal of the protruding nucleotide abated M and L UTR activities and compatibilities with heterogenous viral proteins, and introduction of a protruding nucleotide into S panhandle, conversely, enhanced UTR promoter strength and compatibility, revealing the significance of the protruding nucleotide as a new signature of the genomic panhandle structure in both UTR activity and reassortment potential. The study demonstrates not only banyangvirus reassortment potential but also the notable role of the protruding nucleotide in UTR function and reassortment, providing clues to viral evolution and replication mechanisms and perhaps benefiting disease control and prevention in the future.

中文翻译：

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新兴的Banyangviruses的组合微型基因组系统揭示了病毒重排潜力和基因组“ Panhandle”中突出的核苷酸对于启动子活性和重排的重要性。

Banyangvirus是一个新属（Phenuiviridae家族，Bunyavirales订单），其中包括一组新兴的tick传病毒，伴有血小板减少症候群病毒（SFTSV）和心地病毒（HRTV），表现为严重发烧，是强毒代表。作为分段的RNA病毒，布尼亚病毒可能具有基因组重排的潜力，从而增加了对威胁生命的新布尼亚病毒新出现的担忧。使用基于转染和超级感染的一系列组合微型基因组报告基因检测，我们显示了指定的榕树病毒的复制机制蛋白可以识别其他榕树病毒的基因组非翻译区（UTR），并将异源微型基因组组装为功能性核糖核蛋白（RNP）。此外，病毒糖蛋白将异源和杂合的RNP都有效地包装到了感染性病毒样颗粒中，表现出明显的重组杨病毒的潜力。同时，三个榕树病毒节段的UTR启动子强度似乎是M> L>S。二级结构分析显示，在所有榕树病毒的M和L（而非S）节段的末端UTR Panhandles中保守的非碱基配对突出核苷酸。相关的细小病毒（细小病毒属）。此外，不仅保守的锅柄区域，而且突出的核苷酸对于UTR功能也很重要。去除突出的核苷酸可减弱M和L UTR活性以及与异源病毒蛋白的相容性，并将突出的核苷酸引入S泛柄中，反之，可增强UTR启动子的强度和相容性，揭示了突出核苷酸作为UTR活性和重排潜力的基因组泛柄结构新特征的重要性。这项研究不仅证明了榕树病毒的重排潜力，而且还突出了突出核苷酸在UTR功能和重排中的显著作用，为病毒的进化和复制机制提供了线索，并可能有益于未来的疾病控制和预防。