Ikeda K, Nagaoka S, Winkler S, Kotani K, Yagi H, Nakanishi K, Miyajima S, Kobayashi J, Mori H
Department of Applied Biology, Kyoto Institute of Technology, Kyoto 606-8585, Japan.
J Virol. 2001 Jan;75(2):988-95. doi: 10.1128/JVI.75.2.988-995.2001.
The complete nucleotide sequence of the genome segment 4 (S4) of Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) was determined. The 3,259-nucleotide sequence contains a single long open reading frame which spans nucleotides 14 to 3187 and which is predicted to encode a protein with a molecular mass of about 130 kDa. Western blot analysis showed that S4 encodes BmCPV protein VP3, which is one of the outer components of the BmCPV virion. Sequence analysis of the deduced amino acid sequence of BmCPV VP3 revealed possible sequence homology with proteins from rice ragged stunt virus (RRSV) S2, Nilaparvata lugens reovirus S4, and Fiji disease fijivirus S4. This may suggest that plant reoviruses originated from insect viruses and that RRSV emerged more recently than other plant reoviruses. A chimeric protein consisting of BmCPV VP3 and green fluorescent protein (GFP) was constructed and expressed with BmCPV polyhedrin using a baculovirus expression vector. The VP3-GFP chimera was incorporated into BmCPV polyhedra and released under alkaline conditions. The results indicate that specific interactions occur between BmCPV polyhedrin and VP3 which might facilitate BmCPV virion occlusion into the polyhedra.
家蚕细胞质型多角体病毒(BmCPV)基因组片段4(S4)的完整核苷酸序列已被测定。3259个核苷酸的序列包含一个单一的长开放阅读框,其跨度为核苷酸14至3187,预计编码一种分子量约为130 kDa的蛋白质。蛋白质免疫印迹分析表明,S4编码BmCPV蛋白VP3,它是BmCPV病毒粒子的外部成分之一。对BmCPV VP3推导氨基酸序列的序列分析揭示了与水稻齿叶矮缩病毒(RRSV)S2、褐飞虱呼肠孤病毒S4和斐济病斐济病毒S4的蛋白质可能存在序列同源性。这可能表明植物呼肠孤病毒起源于昆虫病毒,且RRSV比其他植物呼肠孤病毒出现得更晚。构建了一种由BmCPV VP3和绿色荧光蛋白(GFP)组成的嵌合蛋白,并使用杆状病毒表达载体与BmCPV多角体蛋白一起表达。VP3-GFP嵌合体被整合到BmCPV多角体中,并在碱性条件下释放。结果表明,BmCPV多角体蛋白与VP3之间发生了特异性相互作用,这可能有助于BmCPV病毒粒子被包裹进多角体中。