AcMNPV EXON0 (AC141) which is required for the efficient egress of budded virus nucleocapsids interacts with β-tubulin
AcMNPV EXON0 (AC141) which is required for the efficient egress of budded virus nucleocapsids interacts with β-tubulin
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DOI：
10.1016/j.virol.2008.12.023
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发表时间：
2009-03-15
期刊：
影响因子：
3.7
通讯作者：
Theilmann, David A.
中科院分区：
文献类型：
--
作者：
Fang, Minggang;Nie, Yingchao;Theilmann, David A.
苜蓿银纹夜蛾核型多角体病毒（Autographa california multiple nucleopolyhedrovirus，AcMNPV）编码的蛋白EXON 0（AC 141）是将核衣壳有效运输出细胞核以产生出芽病毒（budded virus，BV）所必需的。为了进一步阐明EXON 0调节BV产生的分子机制，用3x FLAG-6x His在N-末端标记EXON 0。通过串联亲和纯化分离蛋白质复合物，并通过LC-MS/MS凝胶纯化和鉴定潜在的EXON 0特异性相互作用蛋白伴侣。该分析表明，细胞蛋白，β-微管蛋白，与EXON 0共纯化，这是通过免疫共沉淀确认的。此外，免疫荧光显示EXON 0和β-微管蛋白在病毒感染期间共定位。用微管抑制剂秋水仙碱和诺考达唑处理AcMNPV感染的Sf 9细胞，结果表明BV的产生减少了85%以上。这些数据表明，AcMNPV出芽病毒的出口可能是由EXON 0与β-微管蛋白和微管的相互作用促进的。皇冠版权所有（c）2008由爱思唯尔公司出版。All rights reserved.
The Autographa california multiple nucleopolyhedrovirus (AcMNPV) encoded protein, EXON0 (AC141), is required for the efficient transport of nucleocapsids out of the nucleus for the production of budded virus (BV). To further elucidate the molecular mechanisms by which EXON0 regulates BV production, EXON0 was tagged at the N-terminus with 3x FLAG-6x His. Protein complexes were isolated by tandem affinity purification and potential EXON0 specific interacting protein partners were gel purified and identified by LC-MS/MS. This analysis showed that the cellular protein, beta-tubulin, co-purified with EXON0 which was confirmed by co-immunoprecipitation. In addition, immunofluorescence showed that EXON0 and beta-tubulin co-localized during Virus infection. The microtubule inhibitors colchicine and nocodazole were used to treat AcMNPV infected Sf9 cells and results showed that BV production was reduced by over 85%. These data suggest that the egress of AcMNPV budded virus may be facilitated by the interaction of EXON0 with beta-tubulin and microtubules. Crown Copyright (c) 2008 Published by Elsevier Inc. All rights reserved.