Wang Hao, Yu Fei, Li Jiale, Lu Liqun
National Pathogen Collection Center for Aquatic Animals, Key Laboratory of Freshwater Fishery Germplasm Resources, Shanghai Ocean University, 201306 Shanghai, PR China.
National Pathogen Collection Center for Aquatic Animals, Key Laboratory of Freshwater Fishery Germplasm Resources, Shanghai Ocean University, 201306 Shanghai, PR China.
Virology. 2016 Mar;490:59-68. doi: 10.1016/j.virol.2016.01.011. Epub 2016 Feb 2.
Grass carp reovirus (GCRV) is responsible for viral hemorrhagic disease in cultured grass carp Ctenopharyngon idellus. Through yeast two-hybrid screen, laminin receptor (LamR) was identified as a potential interacting partner for the outer capsid protein VP5 of GCRV. We cloned and sequenced the gene encoding grass carp LamR. Viral attachment assay demonstrated the involvement of membrane-associated LamR in GCRV infection. Solid-phase overlay assays demonstrated that GCRV interacted with GST-tagged LamR in vitro. In contrast to VP7, GST-tagged VP5 was shown to associate with LamR in both pull-down and solid-phase blot overlay assays. With the reduction of LamR expression in CIK cells achieved by RNAi, remarkably reduced infection efficiency of GCRV was observed. CIK cells pretreated with polyclonal antibody against LamR resulted in dose-dependent inhibition of GCRV infection. These results collectively indicated that grass carp LamR was involved in GCRV infection by interacting with viral outer capsid protein VP5.
草鱼呼肠孤病毒（GCRV）是养殖草鱼（Ctenopharyngon idellus）病毒性出血病的病原体。通过酵母双杂交筛选，层粘连蛋白受体（LamR）被鉴定为GCRV外衣壳蛋白VP5的潜在相互作用伴侣。我们克隆并测序了编码草鱼LamR的基因。病毒附着试验证明膜相关的LamR参与了GCRV感染。固相覆盖试验表明GCRV在体外与GST标记的LamR相互作用。与VP7不同，在下拉试验和固相印迹覆盖试验中，GST标记的VP5均显示与LamR相关。通过RNA干扰降低CIK细胞中LamR的表达后，观察到GCRV的感染效率显著降低。用抗LamR多克隆抗体预处理的CIK细胞导致GCRV感染呈剂量依赖性抑制。这些结果共同表明，草鱼LamR通过与病毒外衣壳蛋白VP5相互作用参与了GCRV感染。
