Non-Structural Protein 5 of Zika Virus Interacts with p53 in Human Neural Progenitor Cells and Induces p53-Mediated Apoptosis
Non-Structural Protein 5 of Zika Virus Interacts with p53 in Human Neural Progenitor Cells and Induces p53-Mediated Apoptosis
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寨卡病毒非结构蛋白 5 与人类神经祖细胞中的 p53 相互作用并诱导 p53 介导的细胞凋亡。
DOI：
10.1007/s12250-021-00422-7
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发表时间：
2021-07-05
影响因子：
5.5
通讯作者：
Li, Chun-Mei
中科院分区：
文献类型：
--
作者：
Li, Ping;Jiang, Hualian;Li, Chun-Mei
寨卡病毒 (ZIKV) 感染可能会通过靶向神经祖细胞 (NPC) 来破坏神经发生并导致新生儿小头畸形。肿瘤抑制因子 p53 介导的细胞周期停滞和细胞凋亡已被认为在 ZIKV 感染后被激活，但详细机制尚不清楚。在本研究中，我们研究了ZIKV编码蛋白在p53信号通路激活中的作用，发现在10种病毒蛋白中，ZIKV的非结构蛋白5（NS5）最显着地激活p53靶基因的转录。使用免疫沉淀偶联质谱方法，我们鉴定出 ZIKV-NS5 与 p53 蛋白相互作用。 NS5-p53 相互作用通过免疫共沉淀和 GST Pull-down 测定进一步得到证实。此外，NS5 的 MTase 结构域和 p53 的 C 末端结构域被映射为负责这两种蛋白质之间的相互作用。我们进一步表明，ZIKV-NS5 与 p53 共定位，增加了其在细胞核中的蛋白水平，并能够延长 p53 的半衰期。此外，hNPC 中慢病毒介导的 ZIKV-NS5 表达导致明显的细胞死亡表型。 ZIKV-NS5 促进 PARP1 的裂解并显着增加 hNPC 的细胞凋亡。总而言之，这些发现表明，ZIKV-NS5 是一种先前未被发现的 hNPC 中 p53 介导的细胞凋亡的调节因子，这可能导致 ZIKV 引起的神经发育异常。
Zika virus (ZIKV) infection could disrupt neurogenesis and cause microcephaly in neonates by targeting neural progenitor cells (NPCs). The tumor suppressor p53-mediated cell cycle arrest and apoptotic cell death have been suggested to be activated upon ZIKV infection, yet the detailed mechanism is not well understood. In the present study, we investigated the effects of ZIKV-encoded proteins in the activation of p53 signaling pathway and found that, among the ten viral proteins, the nonstructural protein 5 (NS5) of ZIKV most significantly activated the transcription of p53 target genes. Using the immunoprecipitation-coupled mass spectrometry approach, we identified that ZIKV-NS5 interacted with p53 protein. The NS5-p53 interaction was further confirmed by co-immunoprecipitation and GST pull-down assays. In addition, the MTase domain of NS5 and the C-terminal domain of p53 were mapped to be responsible for the interaction between these two proteins. We further showed that ZIKV-NS5 was colocalized with p53 and increased its protein level in the nuclei and able to prolong the half-life of p53. Furthermore, lentivirus-mediated expression of ZIKV-NS5 in hNPCs led to an apparent cell death phenotype. ZIKV-NS5 promoted the cleavage of PARP1 and significantly increased the cell apoptosis of hNPCs. Taken together, these findings revealed that ZIKV-NS5 is a previously undiscovered regulator of p53-mediated apoptosis in hNPCs, which may contribute to the ZIKV-caused abnormal neurodevelopment.