Phosphatidylinositol-4 kinase III beta and oxysterol-binding protein accumulate unesterified cholesterol on poliovirus-induced membrane structure
Phosphatidylinositol-4 kinase III beta and oxysterol-binding protein accumulate unesterified cholesterol on poliovirus-induced membrane structure
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DOI：
10.1111/1348-0421.12144
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发表时间：
2014-04-01
影响因子：
2.6
通讯作者：
Arita, Minetaro
中科院分区：
文献类型：
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作者：
Arita, Minetaro
对抗小核糖核酸病毒化合物的研究揭示了通过宿主磷脂酰肌醇-4激酶III β（PI 4KB）和氧固醇结合蛋白（OSBP）家族I的新细胞途径在脊髓灰质炎病毒（PV）复制中的重要作用。然而，该途径在PV复制中的分子作用尚未确定。在这里，病毒和宿主蛋白调节生产的磷脂酰肌醇4-磷酸（PI 4P）和积累的未酯化的胆固醇（UC）在细胞中进行了分析，并在PV复制的PI 4KB/OSBP途径的作用进行了表征。病毒蛋白2BC被鉴定为一种新的PI 4KB相互作用物。PI 4KB和VCP/p97与2BC的部分重叠区域结合，对2C抑制剂具有不同的敏感性。病毒蛋白2BC能促进PI 4P的产生和UC的积累，而病毒蛋白3A和3AB则抑制PI 4P的产生和UC的积累。在PV感染的细胞中，PI 4KB抑制剂抑制PI 4P的产生，并且PI 4KB抑制剂和OSBP配体都抑制UC在病毒诱导的膜结构上的积累。在PV感染的细胞中，PI 4KB活性的抑制导致OSBP从病毒诱导的膜结构中解离。在存在PI 4KB抑制剂和OSBP配体的情况下，PV感染的细胞中病毒新生RNA的合成不受影响;然而，用这些抑制剂对PV感染的细胞进行瞬时预处理抑制了病毒RNA的合成。这些结果表明，病毒蛋白调节PI 4KB活性，并提供PI 4P用于募集OSBP，以在病毒诱导的膜结构上积累UC，从而形成病毒复制复合物。
Studies on anti-picornavirus compounds have revealed an essential role of a novel cellular pathway via host phosphatidylinositol-4 kinase III beta (PI4KB) and oxysterol-binding protein (OSBP) family I in poliovirus (PV) replication. However, the molecular role for this pathway in PV replication has yet to be determined. Here, viral and host proteins modulating production of phosphatidylinositol 4-phosphate (PI4P) and accumulation of unesterified cholesterol (UC) in cells were analyzed and the role of the PI4KB/OSBP pathway in PV replication characterized. Virus protein 2BC was identified as a novel interactant of PI4KB. PI4KB and VCP/p97 bind to a partially overlapped region of 2BC with different sensitivity to a 2C inhibitor. Production of PI4P and accumulation of UC were enhanced by virus protein 2BC, but suppressed by virus proteins 3A and 3AB. In PV-infected cells, a PI4KB inhibitor suppressed production of PI4P, and both a PI4KB inhibitor and an OSBP ligand suppressed accumulation of UC on virus-induced membrane structure. Inhibition of PI4KB activity caused dissociation of OSBP from virus-induced membrane structure in PV-infected cells. Synthesis of viral nascent RNA in PV-infected cells was not affected in the presence of PI4KB inhibitor and OSBP ligand; however, transient pre-treatment of PV-infected cells with these inhibitors suppressed viral RNA synthesis. These results suggest that virus proteins modulate PI4KB activity and provide PI4P for recruitment of OSBP to accumulate UC on virus-induced membrane structure for formation of a virus replication complex.