Expression of measles virus nucleoprotein induces apoptosis and modulates diverse functional proteins in cultured mammalian cells.
Expression of measles virus nucleoprotein induces apoptosis and modulates diverse functional proteins in cultured mammalian cells.
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DOI：
10.1371/journal.pone.0018765
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发表时间：
2011-04-14
期刊：
影响因子：
3.7
通讯作者：
Yadava P
中科院分区：
文献类型：
--
作者：
Bhaskar A;Bala J;Varshney A;Yadava P
麻疹病毒核蛋白(N)封装病毒RNA，保护其免受核酸内切酶的侵害，并形成病毒特异性模板进行转录和复制。它是病毒感染期间最丰富的蛋白质。它的c端结构域本质上是无序的，这赋予了它与几个细胞和病毒伴侣相互作用的灵活性。在这项研究中，我们证明了N在哺乳动物细胞中的表达导致形态学转变、核凝聚、DNA断裂和Caspase 3的激活，最终导致细胞凋亡。细胞内活性氧（ROS）的快速生成参与了细胞死亡的机制。在细胞外培养基中加入抗坏血酸（AA）或caspase-3抑制剂部分逆转N诱导的细胞凋亡。我们还研究了表达N蛋白的细胞的蛋白谱。MS分析显示25个蛋白的差异表达，其中11个蛋白在N表达上上调，14个蛋白在N表达上下调。2DE结果通过实时和半定量RT-PCR分析验证。这些结果显示了N的促凋亡作用，表明其可能发展为凋亡工具。我们的2DE结果提供了初步证据，表明MV核蛋白与多种细胞因子相互作用或导致差异表达。在这个阶段，宿主细胞的适应性反应是什么，以及什么反映了病毒施加的战略调节，还不清楚。
Measles virus nucleoprotein (N) encapsidates the viral RNA, protects it from endonucleases and forms a virus specific template for transcription and replication. It is the most abundant protein during viral infection. Its C-terminal domain is intrinsically disordered imparting it the flexibility to interact with several cellular and viral partners. In this study, we demonstrate that expression of N within mammalian cells resulted in morphological transitions, nuclear condensation, DNA fragmentation and activation of Caspase 3 eventuating into apoptosis. The rapid generation of intracellular reactive oxygen species (ROS) was involved in the mechanism of cell death. Addition of ascorbic acid (AA) or inhibitor of caspase-3 in the extracellular medium partially reversed N induced apoptosis. We also studied the protein profile of cells expressing N protein. MS analysis revealed the differential expression of 25 proteins out of which 11 proteins were up regulated while 14 show signs of down regulation upon N expression. 2DE results were validated by real time and semi quantitative RT-PCR analysis. These results show the pro-apoptotic effects of N indicating its possible development as an apoptogenic tool. Our 2DE results present prima facie evidence that the MV nucleoprotein interacts with or causes differential expression of a wide range of cellular factors. At this stage it is not clear as to what the adaptive response of the host cell is and what reflects a strategic modulation exerted by the virus.
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影响因子：
8
作者：
Bourhis, Jean-Marie;Receveur-Brechot, Veronique;Oglesbee, Michael;Zhang, Xinsheng;Buccellato, Matthew;Darbon, Herve;Canard, Bruno;Finet, Stephanie;Longhi, Sonia
通讯作者：
Longhi, Sonia
影响因子：
14.9
作者：
Jensen LJ;Kuhn M;Stark M;Chaffron S;Creevey C;Muller J;Doerks T;Julien P;Roth A;Simonovic M;Bork P;von Mering C
通讯作者：
von Mering C
影响因子：
5.4
作者：
EGELMAN, EH;WU, SS;MURTI, G
通讯作者：
MURTI, G
影响因子：
4.5
作者：
Muller, Nora;Avota, Elita;Schneider-Schaulies, Sibylle
通讯作者：
Schneider-Schaulies, Sibylle
影响因子：
4.8
作者：
Hishikawa, K;Oemar, BS;Fujii, T
通讯作者：
Fujii, T