ORF005L from infectious spleen and kidney necrosis virus is located in the inner mitochondrial membrane and induces apoptosis
ORF005L from infectious spleen and kidney necrosis virus is located in the inner mitochondrial membrane and induces apoptosis
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DOI：
10.1007/s11262-014-1088-2
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发表时间：
2014-05
期刊：
影响因子：
1.6
通讯作者：
Rui Wang;Y. Yi;Lihong Liu;Yanghong Lu;S. Weng;Jianguo He;Xiaopeng Xu
中科院分区：
文献类型：
--
作者：
Rui Wang;Y. Yi;Lihong Liu;Yanghong Lu;S. Weng;Jianguo He;Xiaopeng Xu
传染性脾肾坏死病毒（infectious spleen and kidney necrosis virus，ISKNV）属于虹彩病毒科巨细胞病毒属（Megalocytivirus）。该病毒是鱼类中一种严重的流行性疾病的病原体。感染ISKNV的细胞发生凋亡。在这项研究中，ISKNV ORF 005 L基因的特点和功能研究。生物信息学分析表明，ORF 005 L蛋白含有一个类似于CTD样磷酸酶催化结构域的区域。实时定量PCR结果显示ORF 005 L在ISKNV感染细胞中的转录谱;在这些细胞中，ORF 005 L在感染后24 h开始转录。绿色荧光蛋白标记的ORF 005 L蛋白定位于线粒体中。Western blot结果显示ORF 005 L特异性定位于线粒体内膜。ORF 005 L在黑头呆鱼细胞中瞬时表达，导致线粒体跨膜电位降低，从而诱导细胞凋亡。ORF 005 L被特异性dsRNA敲低，从而显著降低ISKNV感染诱导的桂鱼fry-1细胞凋亡。这些结果表明，ISKNV的ORF 005 L可以破坏线粒体功能，导致细胞凋亡。本研究为巨细胞病毒感染的发病机制提供了新的认识。
Infectious spleen and kidney necrosis virus (ISKNV) belongs to the genusMegalocytivirusin the family Iridoviridae. This virus is the etiological agent of a serious and pandemic disease in fish. Cells infected with ISKNV undergo apoptosis. In this study, the ISKNV ORF005L gene was characterized and functionally investigated. Bioinformatics analysis revealed that the ORF005L protein contains a region similar to the catalytic domain of CTD-like phosphatases. Real-time quantitative-PCR results showed the transcription profile of ORF005L in ISKNV-infected cells; in these cells, ORF005L was initially transcribed at 24 h post-infection. The green fluorescent protein-tagged ORF005L protein was localized in the mitochondria. Sub-mitochondrial fractions were subjected to Western blot, and the results showed that ORF005L was specifically located in the inner membrane of the mitochondria. The ORF005L in fathead minnow cells was transiently expressed, resulting in the decrease in mitochondrial transmembrane potential, which induced cell apoptosis. ORF005L was knocked down by specific dsRNA, thereby significantly reducing the apoptosis of mandarin fish fry-1 cells induced by ISKNV infection. These results indicated that the ORF005L of ISKNV could disrupt mitochondrial function and cause apoptosis. This study may provide novel insights into the pathogenesis ofMegalocytivirusinfection.