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七肽重复区域对于苜蓿银纹夜蛾核型多角体病毒P10丝的形成至关重要,而富含脯氨酸区域或C末端碱性区域则并非如此。

The heptad repeats region is essential for AcMNPV P10 filament formation and not the proline-rich or the C-terminus basic regions.

作者信息

Dong Chunsheng, Deng Fei, Li Dan, Wang Hualin, Hu Zhihong

机构信息

State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.

出版信息

Virology. 2007 Sep 1;365(2):390-7. doi: 10.1016/j.virol.2007.03.051. Epub 2007 May 3.

Abstract

Baculovirus P10 protein is a small conserved protein and is expressed as bundles of filaments in the host cell during the late phase of virus infection. So far the published results on the domain responsible for filament structural formation have been contradictory. Electron microscopy revealed that the C-terminus basic region was involved in filament structural formation in the Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV) [van Oers, M.M., Flipsen, J.T., Reusken, C.B., Sliwinsky, E.L., Vlak, J.M., 1993. Functional domains of the p10 protein of Autographa californica nuclear polyhedorsis virus. J. Gen. Virol. 74, 563-574.]. While in the Helicoverpa armigera nucleopolyhedrovirus (HearNPV), the heptad repeats region but not the C-terminus domain was proven to be responsible for filament formation [Dong, C., Li, D., Long, G., Deng, F., Wang, H., Hu, Z., 2005. Identification of functional domains required for HearNPV P10 filament formation. Virology 338, 112-120.]. In this manuscript, fluorescence confocal microscopy was applied to study AcMNPV P10 filament formation. A set of plasmids containing different P10 structural domains fused with a fluorescent protein were constructed and transfected into Sf-9 cells. The data indicated that the heptad repeats region, but not the proline-rich region or the C-terminus basic region, is essential for AcMNPV P10 filament formation. Co-transfection of P10s tagged with different fluorescent revealed that P10s with defective heptad repeats region could not interact with intact heptad repeats region or even full-length P10s to form filament structure. Within the heptad repeats region, deletion of the three amino acids spacing of AcMNPV P10 appeared to have no significant impact on the formation of filament structures, but the content of the heptad repeats region appeared to play a role in the morphology of the filaments.

摘要

杆状病毒P10蛋白是一种保守的小蛋白,在病毒感染后期在宿主细胞中以丝状束的形式表达。到目前为止,关于负责丝状结构形成的结构域的已发表结果相互矛盾。电子显微镜显示,在苜蓿银纹夜蛾多核衣壳核多角体病毒(AcMNPV)中,C末端碱性区域参与丝状结构的形成[van Oers, M.M., Flipsen, J.T., Reusken, C.B., Sliwinsky, E.L., Vlak, J.M., 1993.苜蓿银纹夜蛾核多角体病毒p10蛋白的功能结构域。《普通病毒学杂志》74, 563 - 574]。而在棉铃虫核多角体病毒(HearNPV)中,七肽重复区域而非C末端结构域被证明负责丝状结构的形成[Dong, C., Li, D., Long, G., Deng, F., Wang, H., Hu, Z., 2005.棉铃虫核多角体病毒P10丝状结构形成所需功能结构域的鉴定。《病毒学》338, 112 - 120]。在本论文中,应用荧光共聚焦显微镜研究AcMNPV P10丝状结构的形成。构建了一组含有与荧光蛋白融合的不同P10结构域的质粒,并将其转染到Sf - 9细胞中。数据表明,七肽重复区域而非富含脯氨酸区域或C末端碱性区域对于AcMNPV P10丝状结构的形成至关重要。共转染带有不同荧光标签的P10表明,七肽重复区域有缺陷的P10不能与完整的七肽重复区域或甚至全长P10相互作用以形成丝状结构。在七肽重复区域内,删除AcMNPV P10的三个氨基酸间隔似乎对丝状结构的形成没有显著影响,但七肽重复区域的含量似乎在丝状结构的形态中起作用。

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