Liang Yan, Xu Meng-Lin, Wang Xiao-Wen, Gao Xiao-Xiao, Cheng Jun-Jun, Li Chen, Huang Jie
Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, No.106 Nanjing Road, Qingdao, 266071, China.
Virol J. 2015 Mar 29;12:49. doi: 10.1186/s12985-015-0275-7.
Over the past a few years, evidences indicate that adenosine triphosphate (ATP) is an energy source for the binding, maturation, assembly, and budding process of many enveloped viruses. Our previous studies suggest that the F1-ATP synthase beta subunit (ATPsyn β, BP53) of the shrimp Litopenaeus vannamei (L. vannamei) might serve as a potential receptor for white spot syndrome virus (WSSV)'s infection.
BP53 was localized on the surface of shrimp hemocytes and gill epithelial cells by immunofluorescence assay and immunogold labeling technique. Cell surface ATP synthesis was demonstrated by an in vitro bioluminescent luciferase assay. Furthermore, the expression of bp53 after WSSV infection was investigated by RT-PCR test. In addition, RNAi was developed to knock down endogenous bp53.
BP53 is present on shrimp cell surface of hemocytes and gill epithelia. The synthesized ATP was detectable in the extracellular supernatant by using a bioluminescence assay, and the production declined post WSSV binding and infection. Knocking down endogenous bp53 resulted in a 50% mortality of L. vannamei.
These results suggested that BP53, presenting on cell surface, likely served as one of the receptors for WSSV infection in shrimp. Correspondingly, WSSV appears to disturb the host energy metabolism through interacting with host ATPsyn β during infection. This work firstly showed that host ATP production is required and consumed by the WSSV for binding and proceeds with infection process.
在过去几年中,有证据表明三磷酸腺苷(ATP)是许多包膜病毒结合、成熟、组装和出芽过程的能量来源。我们之前的研究表明,凡纳滨对虾(南美白对虾)的F1-ATP合酶β亚基(ATPsynβ,BP53)可能是白斑综合征病毒(WSSV)感染的潜在受体。
通过免疫荧光测定和免疫金标记技术将BP53定位在对虾血细胞和鳃上皮细胞表面。通过体外生物发光荧光素酶测定证明细胞表面ATP合成。此外,通过RT-PCR试验研究WSSV感染后bp53的表达。另外,开发了RNA干扰以敲低内源性bp53。
BP53存在于对虾血细胞和鳃上皮细胞表面。通过生物发光测定法可在细胞外上清液中检测到合成的ATP,并且在WSSV结合和感染后产量下降。敲低内源性bp53导致南美白对虾50%的死亡率。
这些结果表明,存在于细胞表面的BP53可能是对虾中WSSV感染的受体之一。相应地,WSSV在感染过程中似乎通过与宿主ATPsynβ相互作用来干扰宿主能量代谢。这项工作首次表明宿主ATP的产生是WSSV结合和进行感染过程所必需和消耗的。