Junqing Sun, Lu Kuai, Lei Zhang, Yufeng Xie, Yanfang Zhang, Yan Li, Qi Peng, Yuekun Shao, Qiuxian Yang, Wen-Xia Tian, Junhao Zhu, Jianxun Qi, Yi Shi, Tao Deng, George F Gao
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引用次数: 0
摘要
甲型流感病毒(FluA)的基因组转录和复制由病毒 RNA 聚合酶(FluAPol)催化,在整个病毒生命周期中受到微妙的控制。感染后期从转录到复制的转换对于后代病毒的产生至关重要,而病毒非结构蛋白 NS2 被认为参与了这一转换的调控。然而,NS2 的潜在调控机制和结构多年来一直未被发现。在这里,我们以 ~3.0 Å 的分辨率确定了 FluAPol-NS2 复合物的冷冻电镜结构。令人惊讶的是,三个结构域互换的 NS2 二聚体将三个对称的 FluPol 二聚体排列成一个高度有序的桶状六聚体。进一步的结构和功能分析表明,由于立体冲突,NS2 的结合不仅阻碍了 FluAPol 与 Pol II CTD 之间的相互作用,而且还使 FluAPol 在复制酶构象中停滞,从而损害了它的转录酶活性。此外,这是首次可视化的全长 NS2 结构。我们的发现揭示了 FluA 转录-复制转换的关键分子机制,对开发抗病毒药物具有重要意义。
NS2 induces an influenza A RNA polymerase hexamer and acts as a transcription to replication switch.
Genome transcription and replication of influenza A virus (FluA), catalyzed by viral RNA polymerase (FluAPol), are delicately controlled across the virus life cycle. A switch from transcription to replication occurring at later stage of an infection is critical for progeny virion production and viral non-structural protein NS2 has been implicated in regulating the switch. However, the underlying regulatory mechanisms and the structure of NS2 remained elusive for years. Here, we determine the cryo-EM structure of the FluAPol-NS2 complex at ~3.0 Å resolution. Surprisingly, three domain-swapped NS2 dimers arrange three symmetrical FluPol dimers into a highly ordered barrel-like hexamer. Further structural and functional analyses demonstrate that NS2 binding not only hampers the interaction between FluAPol and the Pol II CTD because of steric conflicts, but also impairs FluAPol transcriptase activity by stalling it in the replicase conformation. Moreover, this is the first visualization of the full-length NS2 structure. Our findings uncover key molecular mechanisms of the FluA transcription-replication switch and have implications for the development of antivirals.
期刊介绍:
EMBO Reports is a scientific journal that specializes in publishing research articles in the fields of molecular biology, cell biology, and developmental biology. The journal is known for its commitment to publishing high-quality, impactful research that provides novel physiological and functional insights. These insights are expected to be supported by robust evidence, with independent lines of inquiry validating the findings.
The journal's scope includes both long and short-format papers, catering to different types of research contributions. It values studies that:
Communicate major findings: Articles that report significant discoveries or advancements in the understanding of biological processes at the molecular, cellular, and developmental levels.
Confirm important findings: Research that validates or supports existing knowledge in the field, reinforcing the reliability of previous studies.
Refute prominent claims: Studies that challenge or disprove widely accepted ideas or hypotheses in the biosciences, contributing to the correction and evolution of scientific understanding.
Present null data: Papers that report negative results or findings that do not support a particular hypothesis, which are crucial for the scientific process as they help to refine or redirect research efforts.
EMBO Reports is dedicated to maintaining high standards of scientific rigor and integrity, ensuring that the research it publishes contributes meaningfully to the advancement of knowledge in the life sciences. By covering a broad spectrum of topics and encouraging the publication of both positive and negative results, the journal plays a vital role in promoting a comprehensive and balanced view of scientific inquiry.