Fang Minggang, Nie Yingchao, Dai Xiaojiang, Theilmann David A
Plant Science, Faculty of Land and Food Systems, University of British Columbia, Vancouver, B.C., Canada V6T 1Z4.
Virology. 2008 May 25;375(1):265-76. doi: 10.1016/j.virol.2008.01.036. Epub 2008 Mar 7.
The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) late gene exon0 (ac141) is required for the efficient production of budded virus (BV). EXON0 interacts with nucleopcapsid protein BV/ODV-C42 and FP25 and enables egress of nucleocapsids from the nucleus to the cytoplasm. This study examines the functional domains of EXON0 that play a role in BV production. Six putative domains of the 261 amino acid EXON0 were deleted and examined for functionality by determining their ability to rescue an AcMNPV exon0 knockout bacmid. Domain mapping results showed that all the six domains were required but deletion of the N-terminal acidic region and the leucine zipper domains had the greatest impact on BV production. Yeast 2-hybrid and co-immunoprecipitation demonstrated that EXON0 formed dimers. Point mutation analysis demonstrated that the leucine zipper was required for dimer formation and interaction with BV/ODV-C42 and FP25. The charged domain was also required for BV/ODV-C42 interaction.
苜蓿银纹夜蛾多核衣壳核型多角体病毒（AcMNPV）晚期基因exon0（ac141）是产生出芽病毒（BV）所必需的。EXON0与核衣壳蛋白BV/ODV-C42和FP25相互作用，并使核衣壳从细胞核进入细胞质。本研究检测了EXON0中在BV产生过程中起作用的功能结构域。对261个氨基酸的EXON0的六个假定结构域进行了缺失，并通过测定它们拯救AcMNPV exon0敲除杆粒的能力来检测其功能。结构域定位结果表明，所有六个结构域都是必需的，但N端酸性区域和亮氨酸拉链结构域的缺失对BV产生的影响最大。酵母双杂交和免疫共沉淀表明EXON0形成二聚体。点突变分析表明，亮氨酸拉链是二聚体形成以及与BV/ODV-C42和FP25相互作用所必需的。带电结构域对于BV/ODV-C42的相互作用也是必需的。
