Autographa californica multiple nucleopolyhedrovirus ac53 plays a role in nucleocapsid assembly
Autographa californica multiple nucleopolyhedrovirus ac53 plays a role in nucleocapsid assembly
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苜蓿银纹夜蛾多重核多角体病毒 ac53 在核衣壳组装中发挥作用。
DOI：
10.1016/j.virol.2008.09.003
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发表时间：
2008-12-05
期刊：
影响因子：
3.7
通讯作者：
Pang, Yi
中科院分区：
文献类型：
--
作者：
Liu, Chao;Li, Zhaofei;Pang, Yi
苜蓿银纹夜蛾核型多角体病毒（Autographo californica multiple nucleopolyhedrovirus，AcMNPV）orf 53（ac 53）基因是一个高度保守的基因，存在于所有已测序的鳞翅目和夜蛾目杆状病毒中，但其功能尚不清楚。为了研究其在杆状病毒生命周期中的作用，通过在大肠杆菌中同源重组产生ac 53缺失病毒（vAc（ac 53 KO-PH-GFP））。荧光和光学显微镜和滴定分析表明，vAc（ac 53 KO-PH-GFP）不能在感染的Sf 9细胞中产生感染性出芽病毒。实时荧光定量PCR结果显示ac 53基因缺失对病毒DNA复制水平无影响。电子显微镜显示，许多透明的管状外壳缺乏核蛋白核心存在于病毒发生的基质和环区，表明ac 53基因敲除影响核衣壳组装。利用表达Ac 53-GFP融合蛋白的重组病毒，我们观察到Ac 53在感染后24 h分布在细胞质和细胞核内，但随后主要聚集在细胞核-细胞质边界附近。这些数据表明ac 53参与核衣壳组装，是病毒生产的必需基因。(C)2008年爱思唯尔公司All rights reserved.
Autographo californica multiple nucleopolyhedrovirus (AcMNPV) orf53 (ac53) is a highly conserved gene existing in all sequenced Lepidoptera and Hymenoptera baculoviruses, but its function remains unknown. To investigate its role in the baculovirus life cycle, an ac53 deletion virus (vAc(ac53KO-PH-GFP)) was generated through homologous recombination in Escherichia coli. Fluorescence and light microscopy and titration analysis revealed that vAc(ac53KO-PH-GFP) could not produce infectious budded virus in infected Sf9 cells. Realtime PCR demonstrated that the ac53 deletion did not affect the levels of viral DNA replication. Electron microscopy showed that many lucent tubular shells devoid of the nucleoprotein core are present in the virogenic stroma and ring zone, indicating that the ac53 knockout affected nucleocapsid assembly. With a recombinant virus expressing an Ac53-GFP fusion protein, we observed that Ac53 was distributed within the cytoplasm and nucleus at 24 h post-infection, but afterwards accumulated predominantly near the nucleus-cytoplasm boundary. These data demonstrate that ac53 is involved in nucleocapsid assembly and is an essential gene for virus production. (C) 2008 Elsevier Inc. All rights reserved.