The matrix protein of vesicular stomatitis virus inhibits host-directed transcription of target genes via interaction with the TFIIH subunit p8
The matrix protein of vesicular stomatitis virus inhibits host-directed transcription of target genes via interaction with the TFIIH subunit p8
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水疱性口炎病毒的基质蛋白通过与 TFIIH 亚基 p8 相互作用抑制宿主定向的靶基因转录
DOI：
10.1016/j.vetmic.2017.07.020
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发表时间：
2017-09-01
影响因子：
3.3
通讯作者：
Zhao, Kui
中科院分区：
文献类型：
--
作者：
Pan, Wei;Song, Deguang;Zhao, Kui
响应于病毒感染，引发宿主先天性抗病毒应答以限制病毒复制。许多病毒已经进化出各种策略来规避宿主的抗病毒反应。据报道，水泡性口炎病毒（VSV）的基质（M）蛋白可以抑制宿主基因的表达，从而逃避宿主的天然免疫反应。然而，其分子机制仍不清楚。在这里，我们证明了VSV M蛋白抑制转染到BSR-T7/5细胞中的报告基因的转录。为了进一步研究潜在的机制，进行酵母双杂交筛选以寻找与M蛋白相互作用的宿主蛋白。转录/修复因子TFIIH的亚基p8被鉴定为M结合伴侣，并且用GST下拉试验和激光共聚焦显微镜验证了相互作用。通过突变分析，我们发现当M上的I96、E156、R159和R160残基被Ala取代时，p8-M相互作用受损。这些突变体降低了对报告基因转录的抑制作用。此外，当与p8共表达时，由M介导的转录抑制受损。这些结果表明，p8 M相互作用在抑制宿主基因转录中起重要作用。
In response to viral infection, the host innate antiviral response is elicited to limit viral replication. Many viruses have evolved various strategies to circumvent the host antiviral response. It has been reported that matrix (M) protein of vesicular stomatitis virus (VSV) can inhibit host gene expression to evade the host innate immune response. However, the molecular mechanism remains unclear. Here, we demonstrated that VSV M protein inhibited transcription of a reporter gene transfected into BSR-T7/5 cells. To further investigate the underlying mechanism, a yeast two-hybrid screen was performed to search for host proteins that interact with the M protein. The subunit of transcription/repair factor TFIIH, p8, was identified as an M binding partner, and the interaction was validated with a GST pull-down assay and laser confocal microscopy. Through a mutagenesis analysis, we found that the p8-M interaction was impaired when I96, E156, R159 and R160 residues on M were replaced with Ala. These mutants reduced the inhibitory effect on transcription of the reporter gene. Furthermore, the transcription inhibition mediated by M was impaired when co-expressed with p8. These results indicate that the p8 M interaction plays an important role in inhibiting transcription of host genes.