IJkel W F, Lebbink R J, Op den Brouw M L, Goldbach R W, Vlak J M, Zuidema D
Laboratory of Virology, Wageningen University and Research Centre, Binnenhaven 11, Wageningen, 6709 PD, The Netherlands.
Virology. 2001 Jun 5;284(2):170-81. doi: 10.1006/viro.2001.0906.
Understanding the molecular basis of the distinct biological properties of Spodoptera exigua multicapsid nucleopolyhedrovirus (SeMNPV), such as its narrow host range and high virulence, requires detailed information on the temporal expression and subcellular localization of SeMNPV gene products. The expression of two unique SeMNPV ORFs, 116 (Se116) and 117 (Se117), which show 45% amino acid similarity, was analyzed. Se116 and Se117 were expressed both in cultured cells and in larvae of S. exigua as polyadenylated transcripts of 0.80 and 0.75 kb, respectively. These transcripts initiated from ATCA(G/T)T promoter motifs, commonly found for baculovirus early genes. Se116 transcripts were detected with increasing abundance from 8 to 48 h p.i., whereas Se117 transcripts were present from 4 h p.i. and most abundantly at 24 h p.i. Western blot analysis of infected Se301 cells revealed 27- and 23-kDa proteins for Se116 and Se117, respectively. C-terminal GFP-fusion proteins of Se116 and Se117 were primarily localized in the nucleus of Se301 cells. When Se301 cells were infected with SeMNPV, both GFP-fusion proteins were localized in the virogenic stroma of the nucleus. While the function of the Se116 protein is still enigmatic, the Se117 protein appeared to be a structural protein associated with nucleocapsids of occlusion-derived SeMNPV virions but not of budded virus.
了解甜菜夜蛾多粒包埋核多角体病毒(SeMNPV)独特生物学特性的分子基础,如宿主范围狭窄和高毒力,需要详细了解SeMNPV基因产物的时空表达和亚细胞定位信息。分析了两个独特的SeMNPV开放阅读框116(Se116)和117(Se117)的表达,它们的氨基酸相似性为45%。Se116和Se117在培养细胞和甜菜夜蛾幼虫中均有表达,分别为0.80 kb和0.75 kb的多聚腺苷酸化转录本。这些转录本起始于杆状病毒早期基因常见的ATCA(G/T)T启动子基序。在感染后8至48小时,Se116转录本的丰度逐渐增加,而Se117转录本在感染后4小时出现,并在感染后24小时最为丰富。对感染的Se301细胞进行蛋白质免疫印迹分析显示,Se116和Se117分别有27 kDa和23 kDa的蛋白。Se116和Se117的C端绿色荧光蛋白融合蛋白主要定位于Se301细胞的细胞核中。当Se301细胞被SeMNPV感染时,两种绿色荧光蛋白融合蛋白均定位于细胞核的病毒发生基质中。虽然Se116蛋白的功能仍然不明,但Se117蛋白似乎是一种与源自包涵体的SeMNPV病毒粒子的核衣壳相关的结构蛋白,而与出芽病毒的核衣壳无关。