Function of Actin in Transportation of the AcMNPV from Nuclear to out side of Cell

LI Xiao-qing; ZHOU Rui; JIA Yun-li; JIA Fang; YU Ze-hua; CHEN Xin-wen

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December 2004

LI Xiao-qing, ZHOU Rui, JIA Yun-li, JIA Fang, YU Ze-hua and CHEN Xin-wen. Function of Actin in Transportation of the AcMNPV from Nuclear to out side of Cell[J]. Virologica Sinica, 2004, 19(6): 630-635.

Citation: LI Xiao-qing, ZHOU Rui, JIA Yun-li, JIA Fang, YU Ze-hua, CHEN Xin-wen. Function of Actin in Transportation of the AcMNPV from Nuclear to out side of Cell .VIROLOGICA SINICA, 2004, 19(6) : 630-635.

肌动蛋白在AcMNPV向细胞外运输过程中作用的初步研究*

李小青 ¹ ,
贾蕴丽 ¹ ,
余泽华 ¹ ,
陈新文 ¹ ,

1.
1. 华中师范大学昆虫研究所，湖北武汉 430079
2.
中国科学院武汉病毒研究所，湖北武汉 430071

摘要

本实验利用AcMNPV(Autographa californica multiple nuclear polyhedrosis virus，AcMNPV)的bac-to-bac系统构建了两种重组病毒，即含GFP-actin融合基因的重组病毒AcMNPV-GFP-actin和含GFP基因的重组病毒AcMNPV-GFP。用这两种重组病毒分别感染Sf9细胞，以AcMNPV-GFP感染的Sf9细胞为对照，用共聚焦激光扫描显微镜观察了绿色荧光在病毒感染过程中的分布情况。由于肌动蛋白和绿色荧光蛋白是共定位的，所以绿色荧光的分布情况就是肌动蛋白的分布情况。实验中观察发现，AcMNPV-GFP感染的Sf9细胞中的绿色荧光，在整个感染过程中是弥散分布的，而AcMNPV-GFP-actin感染Sf9细胞后24-72h这段时间内，肌动蛋白最初聚集在细胞核内，随后逐渐由细胞核向细胞质转移，最后完全聚集于细胞膜。根据实验结果，推测肌动蛋白可能参与了AcMNPV出芽型病毒粒子（BV）由细胞核向细胞质运输以及从细胞膜排出的过程。
- 绿色荧光蛋白
- , 肌动蛋白
- , 苜蓿银纹夜蛾多核衣壳核型多角体病毒
- , 重组病毒

Function of Actin in Transportation of the AcMNPV from Nuclear to out side of Cell

LI Xiao-qing ¹ ,
ZHOU Rui ¹ ,
JIA Yun-li ¹ ,
JIA Fang ¹ ,
YU Ze-hua ^1*,, ,
CHEN Xin-wen ²

1.
1. lnstitute of Entomology, Central China Normal University, Wuhan 430079, China
2.
Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China

Corresponding author: YU Ze-hua,

Abstract

To investigate the role of actin in AcMNPV BV transportation from nuclear to cytoplasm, we constructed two recombinant viruses AcMNPV-GFP and AcMNPV-GFP-actin, in which the expression of GFP gene and GFP-actin fusion gene was driven by polyhedrin promoter. Here, we report the expression pattern of GFP in Sf9 cells infected with AcMNPV-GFP and AcMNPV-GFP-actin. Within 24-72 hr after infection, we observed that GFP signal in Sf9 cells infected with AcMNPV-GFP-actin was first concentrated in nuclear and then gradually located to cytoplasm and finally accumulated exclusively at cytoplasmic membrane. In the Sf9 cells infected with AcMNPV-GFP, however, the GFP signal was diffused throughout cell and this expression pattern did not change during the same period of observation. Thus, our results indicated that actin might participate in the transportation of the AcMNPV BV from nuclear to cytoplasm and its exocytosis from the cell.
- Green fluorescent protein
- , Actin
- , AcMNPV
- , Recombinant virus

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Function of Actin in Transportation of the AcMNPV from Nuclear to out side of Cell

Corresponding author: YU Ze-hua,

1. 1. lnstitute of Entomology, Central China Normal University, Wuhan 430079, China
2. Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China

Abstract: To investigate the role of actin in AcMNPV BV transportation from nuclear to cytoplasm, we constructed two recombinant viruses AcMNPV-GFP and AcMNPV-GFP-actin, in which the expression of GFP gene and GFP-actin fusion gene was driven by polyhedrin promoter. Here, we report the expression pattern of GFP in Sf9 cells infected with AcMNPV-GFP and AcMNPV-GFP-actin. Within 24-72 hr after infection, we observed that GFP signal in Sf9 cells infected with AcMNPV-GFP-actin was first concentrated in nuclear and then gradually located to cytoplasm and finally accumulated exclusively at cytoplasmic membrane. In the Sf9 cells infected with AcMNPV-GFP, however, the GFP signal was diffused throughout cell and this expression pattern did not change during the same period of observation. Thus, our results indicated that actin might participate in the transportation of the AcMNPV BV from nuclear to cytoplasm and its exocytosis from the cell.