Heparan Sulfate Proteoglycan Is an Important Attachment Factor for Cell Entry of Akabane and Schmallenberg Viruses
Heparan Sulfate Proteoglycan Is an Important Attachment Factor for Cell Entry of Akabane and Schmallenberg Viruses
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DOI：
10.1128/jvi.00503-17
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发表时间：
2017-05
影响因子：
5.4
通讯作者：
S. Murakami;Akiko Takenaka-Uema;Tomoya Kobayashi;Kentaro Kato;M. Shimojima;M. Palmarini;T. Horimoto
中科院分区：
文献类型：
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作者：
S. Murakami;Akiko Takenaka-Uema;Tomoya Kobayashi;Kentaro Kato;M. Shimojima;M. Palmarini;T. Horimoto
赤羽病毒（Akabane virus，AKAV）和施马伦贝格病毒（Schmallenberg virus，SBV）均为正布尼亚病毒属（Orthobunyavirus）的成员，通过节肢动物媒介传播，具有广泛的细胞嗜性。AKAV和SBV均可引起反刍动物的关节弯曲-积水性无脑综合征。这些正布尼亚病毒进入的主要细胞受体和附着因子尚不清楚。在这里，我们发现，AKAV和SBV感染抑制肝素或酶去除细胞表面硫酸乙酰肝素。为了证实这一发现，我们通过使用成簇规则间隔短回文重复序列（CRISPR）-Cas9系统制备硫酸乙酰肝素蛋白聚糖（HSPG）敲除（KO）细胞，并测量这些病毒与细胞表面的结合量。我们观察到AKAV和SBV与细胞结合的显著减少，限制了这些病毒的感染。这些数据表明，至少在体外，HSPG是AKAV和SBV的重要细胞附着因子，可促进易感细胞中的病毒复制。重要性AKAV和SBV是反刍动物关节弯曲-积水性无脑综合征的病原体，其在畜牧业中造成相当大的经济损失。在这里，我们确定硫酸乙酰肝素蛋白多糖作为AKAV和SBV进入的主要细胞附着因子。此外，我们发现肝素是AKAV和SBV感染的强抑制剂。揭示病毒与宿主相互作用的分子机制对于理解病毒生物学和开发新型减毒活疫苗至关重要。
ABSTRACT Akabane virus (AKAV) and Schmallenberg virus (SBV) are members of the genus Orthobunyavirus, which are transmitted by arthropod vectors with a broad cellular tropism in vitro as well as in vivo. Both AKAV and SBV cause arthrogryposis-hydranencephaly syndrome in ruminants. The main cellular receptor and attachment factor for entry of these orthobunyaviruses are unknown. Here, we found that AKAV and SBV infections were inhibited by the addition of heparin or enzymatic removal of cell surface heparan sulfates. To confirm this finding, we prepared heparan sulfate proteoglycan (HSPG)-knockout (KO) cells by using a clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 system and measured the quantities of binding of these viruses to cell surfaces. We observed a substantial reduction in AKAV and SBV binding to cells, limiting the infections by these viruses. These data demonstrate that HSPGs are important cellular attachment factors for AKAV and SBV, at least in vitro, to promote virus replication in susceptible cells. IMPORTANCE AKAV and SBV are the etiological agents of arthrogryposis-hydranencephaly syndrome in ruminants, which causes considerable economic losses in the livestock industry. Here, we identified heparan sulfate proteoglycan as a major cellular attachment factor for the entry of AKAV and SBV. Moreover, we found that heparin is a strong inhibitor of AKAV and SBV infections. Revealing the molecular mechanisms of virus-host interactions is critical in order to understand virus biology and develop novel live attenuated vaccines.