Involvement of Creatine Kinase B in Hepatitis C Virus Genome Replication through Interaction with the Viral NS4A Protein
Involvement of Creatine Kinase B in Hepatitis C Virus Genome Replication through Interaction with the Viral NS4A Protein
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DOI：
10.1128/jvi.02179-08
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发表时间：
2009-05-15
影响因子：
5.4
通讯作者：
Suzuki, Tetsuro
中科院分区：
文献类型：
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作者：
Hara, Hiromichi;Aizaki, Hideki;Suzuki, Tetsuro
丙型肝炎病毒（HCV）持续感染是慢性肝病的主要原因。本研究的目的是鉴定参与 HCV 复制复合物 (RC) 的宿主细胞因子，并阐明依赖于所鉴定的宿主衍生因子的病毒基因组复制的调节机制。通过对富含 RC 的膜组分进行比较蛋白质组分析以及随后由 RNA 干扰介导的基因沉默，我们确定了参与 HCV 复制的 RC 成分的几种候选成分。我们发现其中一个候选者肌酸激酶 B (CKB) 是一种关键的 ATP 生成酶，可调节非肌肉细胞亚细胞区室中的 ATP，对于 HCV 基因组的有效复制和感染性病毒的传播非常重要。 CKB 与 HCV NS4A 蛋白相互作用，并与 NS3-4A 形成复合物，该复合物具有多种酶活性。 CKB 上调 NS3-4A 介导的 RNA 和 DNA 体外解旋以及透化 HCV 复制细胞中的复制酶活性。我们的结果支持一个模型，其中通过与 NS4A 相互作用将 CKB 招募到 HCV RC 区室（该区室具有高且波动的能量需求）对于病毒基因组的有效复制非常重要。 CKB-NS4A 关联是开发新型抗病毒治疗策略的潜在目标。
Persistent infection with hepatitis C virus (HCV) is a major cause of chronic liver diseases. The aim of this study was to identify host cell factor(s) participating in the HCV replication complex (RC) and to clarify the regulatory mechanisms of viral genome replication dependent on the host-derived factor(s) identified. By comparative proteome analysis of RC-rich membrane fractions and subsequent gene silencing mediated by RNA interference, we identified several candidates for RC components involved in HCV replication. We found that one of these candidates, creatine kinase B (CKB), a key ATP-generating enzyme that regulates ATP in subcellular compartments of nonmuscle cells, is important for efficient replication of the HCV genome and propagation of infectious virus. CKB interacts with HCV NS4A protein and forms a complex with NS3-4A, which possesses multiple enzyme activities. CKB upregulates both NS3-4A-mediated unwinding of RNA and DNA in vitro and replicase activity in permeabilized HCV replicating cells. Our results support a model in which recruitment of CKB to the HCV RC compartment, which has high and fluctuating energy demands, through its interaction with NS4A is important for efficient replication of the viral genome. The CKB-NS4A association is a potential target for the development of a new type of antiviral therapeutic strategy.