Long Noncoding RNA NRAV Promotes Respiratory Syncytial Virus Replication by Targeting the MicroRNA miR-509-3p/RabSc Axis To Regulate Vesicle Transportation
Long Noncoding RNA NRAV Promotes Respiratory Syncytial Virus Replication by Targeting the MicroRNA miR-509-3p/RabSc Axis To Regulate Vesicle Transportation
复制标题
长非编码 RNA NRAV 通过靶向 MicroRNA miR-509-3p/RabSc 轴调节囊泡运输来促进呼吸道合胞病毒复制
DOI：
10.1128/jvi.00113-20
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发表时间：
2020-05-01
影响因子：
5.4
通讯作者：
Wei, Lin
中科院分区：
文献类型：
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作者：
Li, Jian;Li, Miao;Wei, Lin
呼吸道合胞病毒(RSV)是一种囊膜RNA病毒，约占儿童下呼吸道感染的80%。目前的证据支持长非编码RNA(LncRNA)在许多病毒感染性疾病中的功能参与。然而，IncRNAs在RSV感染中的整体生物学效应和临床作用尚不清楚。本研究从lncRNA数据库中获得与呼吸道病毒感染相关的lncRNAs，并收集144份临床痰标本以鉴定与呼吸道病毒感染相关的lncRNAs。定量聚合酶链式反应(QPCR)检测显示，RSV阳性患者的抗病毒应答负性调节因子(NRAV)的表达显著低于未感染患者，而银屑病相关非蛋白编码RNA(PrINS)、核类斑点组装转录本1(NEAT1)和奈托伊沙门氏菌Pas Theler‘s(Nest)在体内和体外的表达无明显差异。同时，NRAV的过表达促进了RSV在A549和BEAS-2B细胞中的增殖，反之亦然，表明NRAV的表达下调是宿主抗病毒防御的一部分。RNA荧光原位杂交(FISH)证实NRAV主要定位于细胞质。通过RNA测序，我们发现囊泡运输蛋白Rab5c的变化趋势与NRAV相同。随后的研究表明，NRAV能够通过海绵microRNA miR-509-3p间接促进RSV的产生，从而释放Rab5c，促进囊泡运输。本研究为病毒与宿主之间通过非编码RNA的相互作用提供了新的视角，可能有助于探索呼吸道病毒潜在的抗病毒靶点。在本研究中，我们发现在RSV感染的患者中，抗病毒应答的长非编码RNA(LncRNA)负性调节因子(NRAV)的表达减少，并且NRAV的过表达在体外促进了RSV的产生，提示RSV感染时NRAV的减少是宿主抗病毒反应的一部分。我们还发现，NRAV与囊泡蛋白Rab5c竞争细胞质中的microRNA miR509-3p，促进RSV的囊泡运输，加速RSV的增殖，从而加深对RSV感染致病机制的理解。
Respiratory syncytial virus (RSV) is an enveloped RNA virus which is responsible for approximately 80% of lower respiratory tract infections in children. Current lines of evidence have supported the functional involvement of long noncoding RNA (lncRNA) in many viral infectious diseases. However, the overall biological effect and clinical role of IncRNAs in RSV infection remain unclear. In this study, lncRNAs related to respiratory virus infection were obtained from the lncRNA database, and we collected 144 clinical sputum specimens to identify lncRNAs related to RSV infection. Quantitative PCR (qPCR) detection indicated that the expression of lncRNA negative regulator of antiviral response (NRAV) in RSV-positive patients was significantly lower than that in uninfected patients, but lncRNA psoriasis-associated non-protein coding RNA induced by stress (PRINS), nuclear paraspeckle assembly transcript 1 (NEAT1), and Nettoie Salmonella pas Theiler's (NeST) showed no difference in vivo and in vitro. Meanwhile, overexpression of NRAV promoted RSV proliferation in A549 and BEAS-2B cells, and vice versa, indicating that the downregulation of NRAV was part of the host antiviral defense. RNA fluorescent in situ hybridization (FISH) confirmed that NRAV was mainly located in the cytoplasm. Through RNA sequencing, we found that Rab5c, which is a vesicle transporting protein, showed the same change trend as NRAV. Subsequent investigation revealed that NRAV was able to favor RSV production indirectly by sponging microRNA miR-509-3p so as to release Rab5c and facilitate vesicle transportation. The study provides a new insight into virus-host interaction through noncoding RNA, which may contribute to exploring potential antivirus targets for respiratory virus.IMPORTANCE The mechanism of interaction between RSV and host noncoding RNAs is not fully understood. In this study, we found that the expression of long noncoding RNA (lncRNA) negative regulator of antiviral response (NRAV) was reduced in RSV-infected patients, and overexpression of NRAV facilitated RSV production in vitro, suggesting that the reduction of NRAV in RSV infection was part of the host antiviral response. We also found that NRAV competed with vesicle protein Rab5c for microRNA miR509-3p in cytoplasm to promote RSV vesicle transport and accelerate RSV proliferation, thereby improving our understanding of the pathogenic mechanism of RSV infection.