Role of La autoantigen and polypyrimidine tract-binding protein in HCV replication
Role of La autoantigen and polypyrimidine tract-binding protein in HCV replication
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DOI：
10.1016/j.virol.2005.02.009
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发表时间：
2005-04-25
期刊：
影响因子：
3.7
通讯作者：
Siddiqui, A
中科院分区：
文献类型：
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作者：
Domitrovich, AM;Diebel, KW;Siddiqui, A
为了确定细胞因子La自身抗原（La）和多聚嘧啶片段结合蛋白（PTB）是否是丙型肝炎病毒（HCV）复制所需的，我们使用siRNA沉默这些因子，然后使用定量RT-PCR监测它们对HCV复制的影响。此外，我们确定了PTB对HCV 3'非编码区（NCR）活性的影响，并研究了其与HCV复制酶复合物组分的相互作用。我们发现，La是有效的HCV复制所必需的，而PTB似乎部分抑制复制。然而，PTB确实阻断HCV RNA依赖性RNA聚合酶（RdRp，NS 5 B）与3 'NCR的结合。间接免疫荧光显微镜显示共定位的细胞质PTB与HCV RdRp在肝癌细胞（Huh-7）表达HCV蛋白，而在体外翻译的病毒蛋白从HCV复制子揭示了相互作用的PTB亚型与NS 5 B聚合酶和NS 3。(c)2005年爱思唯尔公司All rights reserved.
To determine if the cellular factors La autoantigen (La) and polypyrimidine tract-binding protein (PTB) are required for hepatitis C virus (HCV) replication, we used siRNAs to silence these factors and then monitored their effect on HCV replication using quantitative RT-PCR. In addition, we determined the influence of PTB on the activity of the 3' noncoding region (NCR) of HCV and investigated its interaction with the components of the HCV replicase complex. We found that La is essential for efficient HCV replication while PTB appears to partially repress replication. PTB does, however, block the binding of HCV RNA-dependent RNA polymerase (RdRp, NS5B) to the 3'NCR. Indirect immunofluorescence microscopy showed co-localization of cytoplasmic PTB with the HCV RdRp in hepatoma cells (Huh-7) expressing HCV proteins, while in vitro translation of viral proteins from the HCV replicon revealed the interaction of PTB isoforms with NS5B polymerase and NS3. (c) 2005 Elsevier Inc. All rights reserved.