The transmembrane domains of Sindbis virus envelope glycoproteins induce cell death
The transmembrane domains of Sindbis virus envelope glycoproteins induce cell death
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DOI：
10.1128/jvi.72.5.3935-3943.1998
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发表时间：
1998-05-01
影响因子：
5.4
通讯作者：
Levine, B
中科院分区：
文献类型：
--
作者：
Joe, AK;Foo, HH;Levine, B
辛德毕斯病毒是甲病毒的原型，通过诱导细胞凋亡杀死细胞。为了研究辛德毕斯病毒诱导细胞凋亡的潜在机制，我们检测了特定的病毒基因产物是否能够诱导细胞死亡。编码三种结构蛋白衣壳，前体E1（6 K加E1），和前体E2（P62或E3加E2）的基因与β-半乳糖苷酶报告质粒共转染大鼠前列腺腺癌AT 3细胞中的瞬时转染试验。在用6 K加E1和用P62转染的AT 3细胞中观察到细胞死亡，如通过测量蓝色细胞的损失所确定的，但在用衣壳转染的细胞中没有观察到。P62的缺失突变表明大区域的胞质结构域和胞外结构域对于诱导细胞死亡不是必需的。然而，含有与E2跨膜结构域融合的最小E3信号序列和与E1跨膜结构域融合的最小E3信号序列的构建体与全长P62和6 K加E1一样有效地诱导死亡，而用含有与鼠CD 4跨膜结构域连接的E3信号序列的对照构建体转染后没有观察到细胞死亡。这些数据表明辛德毕斯病毒包膜糖蛋白的跨膜结构域的细胞内表达可以杀死AT 3细胞。
Sindbis virus, the prototype alphavirus, kills cells by inducing apoptosis. To investigate potential mechanisms by which Sindbis virus induces apoptosis, we examined whether specific viral gene products were able to induce cell death. Genes encoding the three structural proteins-capsid, the precursor E1 (6K plus E1), and the precursor E2 (P62 or E3 plus E2)-were cotransfected with a beta-galactosidase reporter plasmid in transient-transfection assays in rat prostate adenocarcinoma AT3 cells. Cell death, as determined by measuring the loss of blue cells, was observed in AT3 cells transfected with 6K plus E1 and with P62 but not in cells transfected with capsid, Deletion mutagenesis of P62 indicated that large regions of the cytoplasmic domain and extracellular domain were not essential for the induction of cell death. However, constructs containing the minimal E3 signal sequence fused to the E2 transmembrane domain and the minimal E3 signal sequence fused to the E1 transmembrane domain induced death as efficiently as full-length P62 and 6K plus E1, whereas no cell death was observed after transfection with a control construct containing the E3 signal sequence linked to the transmembrane domain of murine CD4. These data demonstrate that intracellular expression of the transmembrane domains of the Sindbis virus envelope glycoproteins can kill AT3 cells.