Rotavirus activates JNK and p38 signaling pathways in intestinal cells, leading to AP-1-driven transcriptional responses and enhanced virus replication
Rotavirus activates JNK and p38 signaling pathways in intestinal cells, leading to AP-1-driven transcriptional responses and enhanced virus replication
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DOI：
10.1128/jvi.00390-06
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发表时间：
2006-11-01
影响因子：
5.4
通讯作者：
Coulson, Barbara S.
中科院分区：
文献类型：
--
作者：
Holloway, Gavan;Coulson, Barbara S.
已知轮状病毒感染调节许多细胞基因的转录变化。转录因子NF-κ B和AF-1被轮状病毒感染激活，但导致这些事件的上游过程在很大程度上是未知的。因此，我们研究了在轮状病毒感染过程中c-Jun NH 2-末端激酶（JNK）和p38的激活状态，这是已知激活AP-1的激酶。如通过使用磷酸特异性抗体的蛋白质印迹法所评估的，用恒河猴轮状病毒（RRV）感染或暴露于UV-酪蛋白灭活的RRV（I-RRV）导致HT-29、Caco-2和MA 104细胞中JNK的活化。在Caco-2和MA 104细胞中观察到RRV感染期间p38的活化，但在HT-29细胞中未观察到，而暴露于I-RRV并不导致这些细胞系中p38的活化。轮状病毒株SA 11、CRW-8、Wa和UK也激活JNK和p38。与JNK的活化一致，显示AP-1组分c-Jun的磷酸化相应增加。白细胞介素-8（IL-8）和c-jun启动子含有AP-1结合序列，并且这些基因先前已显示在轮状病毒感染期间转录上调。使用JNK（SP 600125）和p38（SB 203580）的特异性抑制剂和实时PCR，我们发现最大的RRV诱导的IL-8和c-jun转录需要JNK和p38活性。这突出了JNK和p38在RRV诱导的AP-1驱动的基因表达中的重要性。值得注意的是，在Caco-2细胞中抑制p38或JNK可降低RRV生长，但不降低病毒结构抗原表达，证明JNK和p38活化对最佳轮状病毒复制的潜在重要性。
Rotavirus infection is known to regulate transcriptional changes in many cellular genes. The transcription factors NF-kappa B and AF-1 are activated by rotavirus infection, but the upstream processes leading to these events are largely unidentified. We therefore studied the activation state during rotavirus infection of c-Jun NH2-terminal kinase (JNK) and p38, which are kinases known to activate AP-1. As assessed by Western blotting using phospho-specific antibodies, infection with rhesus rotavirus (RRV) or exposure to UV-psoralen-inactivated RRV (I-RRV) resulted in the activation of JNK in HT-29, Caco-2, and MA104 cells. Activation of p38 during RRV infection was observed in Caco-2 and MA104 cells but not in HT-29 cells, whereas exposure to I-RRV did not lead to p38 activation in these cell lines. Rotavirus strains SA11, CRW-8, Wa, and UK also activated JNK and p38. Consistent with the activation of JNK, a corresponding increase in the phosphorylation of the AP-1 component c-Jun was shown. The interleukin-8 (IL-8) and c-jun promoters contain AP-1 binding sequences, and these genes have been shown previously to be transcriptionally up-regulated during rotavirus infection. Using specific inhibitors of JNK (SP600125) and p38 (SB203580) and real-time PCR, we showed that maximal RRV-induced IL-8 and c-jun transcription required JNK and p38 activity. This highlights the importance of JNK and p38 in RRV-induced, AP-1-driven gene expression. Significantly, inhibition of p38 or JNK in Caco-2 cells reduced RRV growth but not viral structural antigen expression, demonstrating the potential importance of JNK and p38 activation for optimal rotavirus replication.