Contrasting Roles of Mitogen-Activated Protein Kinases in Cellular Entry and Replication of Hepatitis C Virus: MKNK1 Facilitates Cell Entry
Contrasting Roles of Mitogen-Activated Protein Kinases in Cellular Entry and Replication of Hepatitis C Virus: MKNK1 Facilitates Cell Entry
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DOI：
10.1128/jvi.00954-12
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发表时间：
2013-04-01
影响因子：
5.4
通讯作者：
Lemon, Stanley M.
中科院分区：
文献类型：
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作者：
Kim, Seungtaek;Ishida, Hisashi;Lemon, Stanley M.
人类的亲属组由800多个单独的蛋白激酶组成。它们以多种方式调节细胞新陈代谢，并可能对病毒复制产生直接和间接影响。对于药物开发来说，激酶是诱人的治疗靶点，但对化学抑制剂来说，实现足够的特异性往往是一个挑战。当使用抑制剂来评估c-jun氨基末端(JNK)激酶是否调控丙型肝炎病毒(HCV)复制时，我们遇到了意想不到的非靶点效应，这导致我们发现了一种与丝裂原活化蛋白激酶(MAPK)相关的激酶-MAPK相互作用丝氨酸/苏氨酸激酶1(MKNK1)在病毒进入中的作用。两种JNK抑制剂，AS601245和SP600125，以及RNA干扰(RNAi)介导的JNK1和JNK2的敲除，促进了丙型肝炎病毒复制子RNA的复制，以及将具有感染性的基因组长度的RNA导入Huh-7细胞。JNK基因敲除也增强了感染无细胞病毒后的复制，表明JNK积极地限制了丙型肝炎病毒的复制。尽管如此，AS601245和SP600125都能抑制病毒入侵。筛选一组靶向可能受AS601245和SP600125非靶向效应调节的激酶的抑制剂，使我们确定MKNK1是参与丙型肝炎病毒进入的宿主因子。化学抑制或siRNA敲除MKNK1可显著抑制1a型丙型肝炎病毒和丙型肝炎病毒假型慢病毒颗粒(HCVpp)进入Huh-7细胞，但对病毒RNA复制或细胞增殖和存活的影响很小。我们提出了一个模型，通过该模型，MKNK1可以促进病毒进入表皮生长因子受体(EGFR)和细胞外信号调节激酶(ERK)下游，这两个蛋白都参与了病毒的进入过程。
The human kinome comprises over 800 individual kinases. These contribute in multiple ways to regulation of cellular metabolism and may have direct and indirect effects on virus replication. Kinases are tempting therapeutic targets for drug development, but achieving sufficient specificity is often a challenge for chemical inhibitors. While using inhibitors to assess whether c-Jun N-terminal (JNK) kinases regulate hepatitis C virus (HCV) replication, we encountered unexpected off-target effects that led us to discover a role for a mitogen-activated protein kinase (MAPK)-related kinase, MAPK interacting serine/threonine kinase 1 (MKNK1), in viral entry. Two JNK inhibitors, AS601245 and SP600125, as well as RNA interference (RNAi)-mediated knockdown of JNK1 and JNK2, enhanced replication of HCV replicon RNAs as well as infectious genome-length RNA transfected into Huh-7 cells. JNK knockdown also enhanced replication following infection with cell-free virus, suggesting that JNK actively restricts HCV replication. Despite this, AS601245 and SP600125 both inhibited viral entry. Screening of a panel of inhibitors targeting kinases that may be modulated by off-target effects of AS601245 and SP600125 led us to identify MKNK1 as a host factor involved in HCV entry. Chemical inhibition or siRNA knockdown of MKNK1 significantly impaired entry of genotype 1a HCV and HCV-pseudotyped lentiviral particles (HCVpp) in Huh-7 cells but had only minimal impact on viral RNA replication or cell proliferation and viability. We propose a model by which MKNK1 acts to facilitate viral entry downstream of the epidermal growth factor receptor (EGFR) and extracellular signal-regulated kinase (ERK), both of which have been implicated in the entry process.