Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands.
Netherlands Cancer Institute, Amsterdam, The Netherlands.
J Virol. 2019 Jun 14;93(13). doi: 10.1128/JVI.02160-18. Print 2019 Jul 1.
Vaccinia virus is a promising viral vaccine and gene delivery candidate and has historically been used as a model to study poxvirus-host cell interactions. We employed a genome-wide insertional mutagenesis approach in human haploid cells to identify host factors crucial for vaccinia virus infection. A library of mutagenized HAP1 cells was exposed to modified vaccinia virus Ankara (MVA). Deep-sequencing analysis of virus-resistant cells identified host factors involved in heparan sulfate synthesis, Golgi organization, and vesicular protein trafficking. We validated EXT1, TM9SF2, and TMED10 (TMP21/p23/p24δ) as important host factors for vaccinia virus infection. The critical roles of EXT1 in heparan sulfate synthesis and vaccinia virus infection were confirmed. TM9SF2 was validated as a player mediating heparan sulfate expression, explaining its contribution to vaccinia virus infection. In addition, TMED10 was found to be crucial for virus-induced plasma membrane blebbing and phosphatidylserine-induced macropinocytosis, presumably by regulating the cell surface expression of the TAM receptor Axl. Poxviruses are large DNA viruses that can infect a wide range of host species. A number of these viruses are clinically important to humans, including variola virus (smallpox) and vaccinia virus. Since the eradication of smallpox, zoonotic infections with monkeypox virus and cowpox virus are emerging. Additionally, poxviruses can be engineered to specifically target cancer cells and are used as a vaccine vector against tuberculosis, influenza, and coronaviruses. Poxviruses rely on host factors for most stages of their life cycle, including attachment to the cell and entry. These host factors are crucial for virus infectivity and host cell tropism. We used a genome-wide knockout library of host cells to identify host factors necessary for vaccinia virus infection. We confirm a dominant role for heparin sulfate in mediating virus attachment. Additionally, we show that TMED10, previously not implicated in virus infections, facilitates virus uptake by modulating the cellular response to phosphatidylserine.
痘病毒是一种很有前途的病毒疫苗和基因传递候选物，历史上一直被用作研究痘病毒-宿主细胞相互作用的模型。我们在人单倍体细胞中采用全基因组插入诱变方法，鉴定对痘病毒感染至关重要的宿主因子。将突变的 HAP1 细胞文库暴露于改良的安卡拉痘苗病毒（MVA）中。对抗病毒细胞的深度测序分析鉴定了参与硫酸乙酰肝素合成、高尔基体组织和囊泡蛋白运输的宿主因子。我们验证了 EXT1、TM9SF2 和 TMED10（TMP21/p23/p24δ）是痘病毒感染的重要宿主因子。EXT1 在硫酸乙酰肝素合成和痘病毒感染中的关键作用得到了证实。TM9SF2 被验证为介导硫酸乙酰肝素表达的参与者，解释了其对痘病毒感染的贡献。此外，发现 TMED10 对于病毒诱导的质膜起泡和磷脂酰丝氨酸诱导的巨胞饮作用至关重要，这可能是通过调节 TAM 受体 Axl 的细胞表面表达来实现的。痘病毒是一种可以感染多种宿主物种的大型 DNA 病毒。其中一些病毒对人类具有重要的临床意义，包括天花病毒（天花）和痘苗病毒。自天花根除以来，猴痘病毒和牛痘病毒的人畜共患感染正在出现。此外，痘病毒可以被工程化以专门针对癌细胞，并被用作针对结核病、流感和冠状病毒的疫苗载体。痘病毒在其生命周期的大多数阶段都依赖宿主因子，包括与细胞的附着和进入。这些宿主因子对病毒的感染力和宿主细胞的嗜性至关重要。我们使用宿主细胞的全基因组敲除文库来鉴定痘病毒感染所必需的宿主因子。我们证实肝素硫酸酯在介导病毒附着中起主要作用。此外，我们表明，TMED10 以前未被认为与病毒感染有关，通过调节对磷脂酰丝氨酸的细胞反应，促进病毒摄取。
