Ac MNPV ie-1基因诱导的斜纹夜蛾细胞凋亡(英文)  

Apoptosis ofSpodoptera litura Cells Induced by AcMNPVie-1Gene

作　　者：张萍[1] 杨波[1] 代小江[1] 庞义[1] 钟江[2] 苏德明[2] 

机构地区：[1]中山大学生物防治国家重点实验室,广州510275 [2]复旦大学生命科学学院病毒学研究室,上海200433

出　　处：《生物化学与生物物理学报》2002年第6期707-711,共5页

基　　金：国家自然科学基金资助项目 (No .39730030,No .3980 0092 )~~

摘　　要：苜蓿丫纹夜蛾核多角体病毒 (Autographacalifornicamulticapsidnucleopolyhedrovirus,AcMNPV)感染可诱导斜纹夜蛾 (Spodopteralitura)离体细胞Sl zsu 1发生典型的细胞凋亡。通过细胞松弛素 (cytochalasinD)和NH4Cl的抑制实验 ,分别排除病毒粒子结合细胞受体蛋白 ,和病毒在核内体运输过程启动细胞凋亡信号发生的可能性。RT PCR实验证实 ,病毒基因组进入了细胞核 ,极早期基因ie 1开始了转录 ;而DNA聚合酶抑制剂 (芽栖菌素 )的存在对病毒诱导的细胞凋亡程度与进程均没有明显的影响。这说明细胞凋亡的信号是先于病毒晚期复制事件启动的。单独转染AcMNPV极早期基因ie 1可诱导斜纹夜蛾离体细胞系Sl zsu 1细胞发生部分凋亡 ,转染 2 4h后出现凋亡小体 ,4 8h达到高峰。提取转染细胞的总DNA电泳 ,可检测到典型的DNA梯形条带 (DNAladder)。另外 ,AcMNPV的ie 1基因温度敏感突变株tsB82 1在非受纳温度感染细胞时 ,细胞不发生凋亡。这些结果暗示 ,在AcMNPV感染诱导的Sl zsu 1细胞凋亡中 ,ie 1基因是一个凋亡信号的直接或间接诱导因子。Apoptosis was induced inSpodoptera lituraSl-zsu-1 cells transfected with plasmid pAcie-1 DNA containingie-1 gene ofAutographa californica multicapsid nucleopolyhedrovirus (AcMNPV). Apoptotic bodies appeared 24 h post transfection and cell mortality peaked after 48 h. Electrophoresis of total DNA extract of Sl-zsu-1 cells showed a DNA ladder, indicating that transfection ofie-1 gene alone can trigger apoptosis directly or indirectly. It was shown by cytochalasin D (CD) and ammonium chloride inhibition assays that the entry of virions into cytoplasm was required for induction of apoptosis. Reverse transcriptase PCR (RT-PCR) was used to demonstrate thatie-1 gene had transcribed in Sl-zsu-1 cells immediately after infection. Inhibition of DNA replication could not block the cell death process, suggesting that this apoptosis was independent on viral late replication events. Apoptosis did not occur when Sl-zsu-1 cells were infected with an AcMNPV mutant tsB821 at its nonpermissive temperature. Therefore, we concluded that theie-1 gene is at least one of the factors inducing apoptosis in Sl-zsu-1 cells infected with AcMNPV.

关 键 词：AcMNPVie-1基因 诱导 斜纹夜蛾 细胞凋亡 

分 类 号：Q753[生物学—分子生物学]