PML tumor suppressor protein is required for HCV production
PML tumor suppressor protein is required for HCV production
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DOI：
10.1016/j.bbrc.2012.11.108
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发表时间：
2013-01-11
影响因子：
3.1
通讯作者：
Kato, Nobuyuki
中科院分区：
文献类型：
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作者：
Kuroki, Misao;Ariumi, Yasuo;Kato, Nobuyuki
PML肿瘤抑制蛋白形成离散的核结构，称为PML核体，与多种细胞功能有关，包括细胞增殖、凋亡和抗病毒防御。最近有报道称，在PML-NBS中，丙型肝炎病毒核心蛋白与PML共存，并通过与PML相互作用而取消PML功能。然而，PML在丙型肝炎病毒生命周期中的作用(S)尚不清楚。为了验证PML是否影响丙型肝炎病毒的生命周期，我们检测了培养上清液中丙型肝炎病毒核心蛋白的分泌水平、丙型肝炎病毒的感染性以及在HU-7来源的RSC细胞中丙型肝炎病毒RNA的水平，在该细胞中，丙型肝炎病毒JFH1可以感染并高效复制，稳定表达针对PML的短发夹状RNA。在此背景下，PML基因敲除细胞分泌的丙型肝炎病毒核心水平和培养上清液中的感染性显著降低，而尽管PML基因敲除非常有效，但PML基因敲除细胞中的HCVRNA水平并未受到显著影响。事实上，我们使用亚基因组丙型肝炎病毒-JFH1复制子RNA JRN/3-5B证明了PML与丙型肝炎病毒RNA复制无关。此外，在表达丙型肝炎病毒JFH1基因核心到NS2编码区的PML敲除的JRN/3-5B细胞中，培养上清液中的丙型肝炎病毒样颗粒的感染力显著降低。最后，我们还证明了PML相关蛋白INI1和DDX5参与了丙型肝炎病毒的产生。综上所述，这些发现表明PML是丙型肝炎病毒产生所必需的。皇冠版权所有(C)2012由爱思唯尔公司出版。保留所有权利。
PML tumor suppressor protein, which forms discrete nuclear structures termed PML-nuclear bodies, has been associated with several cellular functions, including cell proliferation, apoptosis and antiviral defense. Recently, it was reported that the HCV core protein colocalizes with PML in PML-NBs and abrogates the PML function through interaction with PML. However, role(s) of PML in HCV life cycle is unknown. To test whether or not PML affects HCV life cycle, we examined the level of secreted HCV core and the infectivity of HCV in the culture supernatants as well as the level of HCV RNA in HuH-7-derived RSc cells, in which HCV-JFH1 can infect and efficiently replicate, stably expressing short hairpin RNA targeted to PML. In this context, the level of secreted HCV core and the infectivity in the supernatants from PML knockdown cells was remarkably reduced, whereas the level of HCV RNA in the PML knockdown cells was not significantly affected in spite of very effective knockdown of PML. In fact, we showed that PML is unrelated to HCV RNA replication using the subgenomic HCV-JFH1 replicon RNA, JRN/3-5B. Furthermore, the infectivity of HCV-like particle in the culture supernatants was significantly reduced in PML knockdown JRN/3-5B cells expressing core to NS2 coding region of HCV-JFH1 genome using the trans-packaging system. Finally, we also demonstrated that INI1 and DDX5, the PML-related proteins, are involved in HCV production. Taken together, these findings suggest that PML is required for HCV production. Crown Copyright (C) 2012 Published by Elsevier Inc. All rights reserved.