Hantaviruses are causing life-threatening zoonotic infections in humans. Their tripartite negative-stranded RNA genome is replicated by the multi-functional viral RNA-dependent RNA-polymerase. Here we describe the structure of the Hantaan virus polymerase core and establish conditions for in vitro replication activity. The apo structure adopts an inactive conformation that involves substantial folding rearrangement of polymerase motifs. Binding of the 5′ viral RNA promoter triggers Hantaan virus polymerase reorganization and activation. It induces the recruitment of the 3′ viral RNA towards the polymerase active site for prime-and-realign initiation. The elongation structure reveals the formation of a template/product duplex in the active site cavity concomitant with polymerase core widening and the opening of a 3′ viral RNA secondary binding site. Altogether, these elements reveal the molecular specificities of Hantaviridae polymerase structure and uncover the mechanisms underlying replication. They provide a solid framework for future development of antivirals against this group of emerging pathogens.

汉坦病毒正在人类中引起危及生命的人畜共患感染。它们的三重负链 RNA 基因组由多功能病毒 RNA 依赖性 RNA 聚合酶复制。在这里，我们描述了汉坦病毒聚合酶核心的结构，并建立了体外复制活性的条件。apo 结构采用非活性构象，涉及聚合酶基序的大量折叠重排。5' 病毒 RNA 启动子的结合触发汉坦病毒聚合酶重组和激活。它诱导 3' 病毒 RNA 向聚合酶活性位点募集，用于启动和重新对齐启动。延伸结构揭示了活性位点空腔中模板/产物双链体的形成，伴随着聚合酶核心加宽和 3' 病毒 RNA 二级结合位点的开放。汉坦病毒科聚合酶结构和揭示复制的机制。它们为未来开发针对这组新兴病原体的抗病毒药物提供了坚实的框架。