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比较转录组分析揭示 WSSV IE1 蛋白在 DNA 复制控制中起着关键作用。

Comparative Transcriptome Analysis Reveals That WSSV IE1 Protein Plays a Crucial Role in DNA Replication Control.

机构信息

Institute of Marine Sciences and Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China.

出版信息

Int J Mol Sci. 2022 Jul 25;23(15):8176. doi: 10.3390/ijms23158176.

Abstract

For DNA viruses, the immediate-early (IE) proteins are generally essential regulators that manipulate the host machinery to support viral replication. Recently, IE1, an IE protein encoded by white spot syndrome virus (WSSV), has been demonstrated to function as a transcription factor. However, the target genes of IE1 during viral infection remain poorly understood. Here, we explored the host target genes of IE1 using RNAi coupled with transcriptome sequencing analysis. A total of 429 differentially expressed genes (DEGs) were identified from penaeid shrimp, of which 284 genes were upregulated and 145 genes were downregulated after IE1 knockdown. GO and KEGG pathway enrichment analysis revealed the identified DEGs are significantly enriched in the minichromosome maintenance (MCM) complex and DNA replication, indicating that IE1 plays a critical role in DNA replication control. In addition, it was found that MCM complex genes were remarkably upregulated after WSSV infection, while RNAi-mediated knockdown of MCM2 reduced the expression of viral genes and viral loads at the early infection stage. Finally, we demonstrated that overexpression of IE1 promoted the expression of MCM complex genes as well as cellular DNA synthesis in insect High-Five cells. Collectively, our current data suggest that the WSSV IE1 protein is a viral effector that modulates the host DNA replication machinery for viral replication.

摘要

对于 DNA 病毒,早期(IE)蛋白通常是关键的调节因子,它们操纵宿主机制以支持病毒复制。最近,已证实白斑综合征病毒(WSSV)编码的 IE1 蛋白作为转录因子发挥作用。然而,IE1 在病毒感染过程中的靶基因仍知之甚少。在这里,我们使用 RNAi 结合转录组测序分析来探索 IE1 的宿主靶基因。从对虾中鉴定出 429 个差异表达基因(DEGs),IE1 敲低后其中 284 个基因上调,145 个基因下调。GO 和 KEGG 通路富集分析显示,鉴定出的 DEGs 显著富集于微小染色体维持(MCM)复合物和 DNA 复制途径,表明 IE1 在 DNA 复制控制中发挥关键作用。此外,发现 WSSV 感染后 MCM 复合物基因显著上调,而 MCM2 的 RNAi 介导敲低降低了早期感染阶段病毒基因的表达和病毒载量。最后,我们证明 IE1 的过表达促进了昆虫 High-Five 细胞中 MCM 复合物基因的表达和细胞 DNA 合成。总之,我们目前的数据表明,WSSV IE1 蛋白是一种病毒效应物,可调节宿主 DNA 复制机制以促进病毒复制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8e2d/9330391/c2838443a92a/ijms-23-08176-g001.jpg

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