DEFA1B inhibits ZIKV replication and retards cell cycle progression through interaction with ORC1.
DEFA1B inhibits ZIKV replication and retards cell cycle progression through interaction with ORC1.
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DOI：
10.1016/j.lfs.2020.118564
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发表时间：
2020-12-15
期刊：
影响因子：
6.1
通讯作者：
Chen L
中科院分区：
文献类型：
--
作者：
Li S;Zhu A;Ren K;Li S;Chen L
寨卡病毒（ZIKV）感染引起了公共卫生问题，因为它与小头畸形的发展有潜在的关联。在病毒感染过程中，宿主的先天免疫反应迅速启动，产生一些内源性功能分子，限制病毒的复制和传播。外来体含有来自病毒感染后其来源细胞的分子，并且可以进入受体细胞进行细胞间通讯。在这里，我们的目标是澄清ZIKV诱导的外泌体是否可以通过转移特定的RNA来调节病毒的致病性。在该研究中，从具有或不具有ZIKV感染的A549细胞的上清液中分离外来体。进行人转录组阵列（HTA）以分析包裹在外来体中的RNA的谱。然后使用qPCR、蛋白质印迹和ELISA来确定ZIKV复制。CCK-8法和流式细胞仪检测细胞增殖和细胞周期。采用共培养法分析exosomes对受体细胞周期的影响。通过人类转录组阵列（HTA），我们发现从ZIKV感染的A549细胞中分离的外泌体内防御素α 1B（DEFA 1B）表达显著增加。此外，我们发现细胞外DEFA 1B主要在ZIKV进入宿主细胞之前发挥显著的抗ZIKV活性。有趣的是，上调的DEFA 1B会阻滞宿主细胞的细胞周期。进一步的研究表明，DEFA 1B与细胞周期中启动DNA复制所需的起始识别复合物1（ORC 1）相互作用，并且DEFA 1B表达的增加降低了细胞核中ORC 1的水平。因此，含有DEFA 1B的外泌体可以被受体细胞内化以延迟它们的细胞周期。总之，我们的结果表明DEFA 1B的抗ZIKV活性可以由外来体介导，并且DEFA 1B与ORC 1相互作用以延迟细胞周期。我们的研究提供了一个新的概念，即DEFA 1B不仅在ZIKV感染期间充当抗病毒分子，而且还可能通过延缓细胞周期的进展与细胞增殖相关。
Zika virus (ZIKV) infection causes a public health concern because of its potential association with the development of microcephaly. During viral infections, the host innate immune response is mounted quickly to produce some endogenous functional molecules to limit virus replication and spread. Exosomes contain molecules from their cell of origin following virus infection and can enter recipient cells for intercellular communication. Here, we aim to clarify whether ZIKV-induced exosomes can regulate viral pathogenicity by transferring specific RNAs. In this study, exosomes were isolated from the supernatants of A549 cells with or without ZIKV infection. Human transcriptome array (HTA) was performed to analyze the profiling of RNAs wrapped in exosomes. Then qPCR, western blotting and ELISA were used to determine ZIKV replication. CCK-8 and flow cytometry were used to test the cell proliferation and cell cycles. Co-culture assay was used to analyze the effect of exosomes on the cell cycles of recipient cells. Through human transcriptome array (HTA) we found the defensin alpha 1B (DEFA1B) expression was significantly increased within exosomes isolated from ZIKV infected A549 cells. Additionally, we found that the extracellular DEFA1B exerts significant anti-ZIKV activity, mainly before ZIKV entering host cells. Interestingly, up-regulated DEFA1B retards the cell cycle of host cells. Further studies demonstrated that DEFA1B interacted with the origin recognition complex 1 (ORC1) which is required to initiate DNA replication during the cell cycle and increased DEFA1B expression decreased the ORC1 level in the cell nuclei. Accordingly, DEFA1B-containing exosomes can be internalized by the recipient cells to retard their cell cycles. Together, our results demonstrated that the anti-ZIKV activity of DEFA1B can be mediated by exosomes, and DEFA1B interacts with ORC1 to retard cell cycles. Our study provides a novel concept that DEFA1B not only acts as an antiviral molecule during ZIKV infection but also may correlate with cell proliferation by retarding the progression of cell cycles.
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