Identification of hnRNPH1, NF45, and C14orf166 as Novel Host Interacting Partners of the Mature Hepatitis C Virus Core Protein
Identification of hnRNPH1, NF45, and C14orf166 as Novel Host Interacting Partners of the Mature Hepatitis C Virus Core Protein
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DOI：
10.1021/pr200338d
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发表时间：
2011-10-01
影响因子：
4.4
通讯作者：
Wang, Shainn-Wei
中科院分区：
文献类型：
--
作者：
Lee, Jun-Wei;Liao, Pao-Chi;Wang, Shainn-Wei
丙型肝炎病毒核心蛋白（HCVc）形成病毒核衣壳，并参与病毒的持久性和发病机制，可能通过与宿主因子相互作用来调节病毒复制和细胞功能。在这里，我们确定了36个细胞蛋白候选人的一维SDS-PAGE和LC-MS/MS为基础的蛋白质组学与HCVc 174，HCVc的成熟形式从HCV-1b基因型，标记生物素和钙调蛋白结合肽/蛋白A的N-和C-末端，分别亲和纯化后。通过下拉和共聚焦成像技术，我们证实，异质核核糖核蛋白H1（hnRNPH 1），核因子45（NF 45），和C14 orf 166是新的HCVc 174相互作用的宿主蛋白，已知参与mRNA代谢，基因调控和微管组织，分别。与其他两种蛋白不同，NF 45与HCVc 174以RNA依赖的方式相互作用。这3个蛋白与异位HCVc-1b共定位在细胞质和细胞核中，这表明它们与HCVc生物合成后自然易位的HCVc 174的空间相互作用。然而，这种共定位转移到细胞质中的复制病毒的1b或2a基因型，表明活跃的病毒复制局限于这些相互作用的蛋白质在细胞质中。总的来说，我们的研究结果表明，hnRNPH 1，NF 45和C14 orf 166与HCVc 174的空间相互作用可能在急性和慢性HCV感染期间调节HCV或细胞功能。
The hepatitis C virus core protein (HCVc) forms the viral nucleocapsid and is involved in viral persistence and pathogenesis, possibly by interacting with host factors to modulate viral replication and cellular functions. Here, we identified 36 cellular protein candidates by one-dimensional SDS-PAGE and LC-MS/MS-based proteomics after affinity purification with HCVc174, a matured form of HCVc from HCV-1b genotype, tagged with biotin and calmodulin-binding peptide/protein A at N- and C-termini, respectively. By pull-down and confocal imaging techniques, we confirmed that heterogeneous nuclear ribonucleoprotein H1 (hnRNPH1), nuclear factor 45 (NF45), and C14orf166 are novel HCVc174-interacting host proteins, known to participate in mRNA metabolism, gene regulation, and microtubule organization, respectively. Unlike the other 2 proteins, NF45 interacted with HCVc174 in an RNA-dependent manner. These 3 proteins colocalized with ectopic HCVc-1b in both the cytoplasm and nucleus, which demonstrated their spatial interaction with naturally translocated HCVc174 after HCVc biogenesis. Such colocalization, however, shifted to the cytoplasm in cells with replicating virus of 1b or 2a genotype, indicating that active viral replication confined these interacting proteins in the cytoplasm. Collectively, our findings suggest that spatial interactions of hnRNPH1, NF45, and C14orf166 with HCVc174 likely modulate HCV or cellular functions during acute and chronic HCV infection.