近日，國際學(xué)術(shù)期刊《PANS》在線(xiàn)發(fā)表了病毒學(xué)國家重點(diǎn)實(shí)驗室崔宗強教授團隊的最新研究成果，論文題為Real-time dissection of dynamic uncoating of individual influenzaviruses（實(shí)時(shí)動(dòng)態(tài)解析單個(gè)流感病毒脫殼過(guò)程）。該團隊利用量子點(diǎn)特異性標記病毒的基因組和單顆粒示蹤技術(shù)，實(shí)時(shí)動(dòng)態(tài)解析了單個(gè)流感病毒的脫殼過(guò)程，揭示了病毒八節段RNA以單體形式釋放到細胞質(zhì)并進(jìn)入細胞核及其在細胞核內的動(dòng)態(tài)行為及機制。

結果速覽

該團隊首先建立了量子點(diǎn)位點(diǎn)特異性標記甲型流感病毒基因組技術(shù)。在病毒感染過(guò)程中，病毒核糖核蛋白復合物（vRNP）頂端的聚合酶蛋白PA被可控生物素化，與鏈霉親和素修飾的量子點(diǎn)特異性結合，并組裝進(jìn)入成熟的子代病毒粒子，從而獲得基因組被量子點(diǎn)標記的流感病毒?；诹孔狱c(diǎn)的高亮度和耐光漂白等特點(diǎn)，利用單顆粒示蹤技術(shù)，結合病毒包膜和細胞組分熒光標記，對單顆流感病毒感染宿主細胞時(shí)的脫殼過(guò)程進(jìn)行實(shí)時(shí)動(dòng)態(tài)可視化分析。實(shí)時(shí)動(dòng)態(tài)揭示了流感病毒在感染后30-90分鐘，病毒由早期內體進(jìn)入晚期內體發(fā)生膜融合、基因組與病毒包膜分離、單個(gè)節段基因從晚期內體中分別釋放、解離釋放的單個(gè)節段基因入核、及其在細胞核內運輸的動(dòng)態(tài)過(guò)程。發(fā)現流感病毒八個(gè)vRNP在脫殼過(guò)程中以單獨而非組團的方式從晚期內體中釋放進(jìn)入細胞質(zhì)，這些單獨的vRNP以特征性三階段主動(dòng)運輸機制進(jìn)入細胞核，入核后的vRNP以?xún)煞N不同的擴散模式運輸到其復制/轉錄位點(diǎn)等重要過(guò)程與機制。該研究為世界上首次記錄到單個(gè)流感病毒脫殼的實(shí)時(shí)動(dòng)態(tài)過(guò)程，對深入理解流感病毒在宿主細胞中的生命周期具有重要意義，并將為開(kāi)發(fā)新的抗病毒途徑提供思路。

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ABSTRACT: Uncoating is an obligatory step in the virus life cycle that serves as an antiviral target. Unfortunately, it is challenging to study viral uncoating due to methodology limitations for detecting this transient and dynamic event. The uncoating of influenza A virus (IAV), which contains an unusual genome of eight segmented RNAs, is particularly poorly understood. Here, by encapsulating quantum dot (QD)-conjugated viral ribonucleoprotein complexes (vRNPs) within infectious IAV virions and applying single-particle imaging, we tracked the uncoating process of individual IAV virions. Approximately 30% of IAV particles were found to undergo uncoating through fusion with late endosomes in the “around-nucleus” region at 30 to 90 minutes postinfection. Inhibition of viral M2 proton channels and cellular endosome acidification prevented IAV uncoating. IAV vRNPs are released separately into the cytosol after virus uncoating. Then, individual vRNPs undergo a three-stage movement to the cell nucleus and display two diffusion patterns when inside the nucleus. These findings reveal IAV uncoating and vRNP trafficking mechanisms, filling a critical gap in knowledge about influenza viral infection.

中國科學(xué)院武漢病毒研究所崔宗強研究員為該論文通訊作者，秦沖博士為第一作者。該研究得到了中國科學(xué)院戰略性先導科技專(zhuān)項、國家重點(diǎn)研發(fā)計劃、國家自然科學(xué)基金、中國科學(xué)院青年創(chuàng )新促進(jìn)會(huì )等基金項目的資助。

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參考：

1.原文鏈接：

https://doi.org/10.1073/pnas.1812632116

2.病毒學(xué)國家重點(diǎn)實(shí)驗室官網(wǎng)：

https://klv.whu.edu.cn/index.php/View/321.html
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文章內容來(lái)源：病毒學(xué)界

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