Hidden regulation of herpes simplex virus 1 pre-mRNA splicing and polyadenylation by virally encoded immediate early gene ICP27
Hidden regulation of herpes simplex virus 1 pre-mRNA splicing and polyadenylation by virally encoded immediate early gene ICP27
复制标题
病毒编码的立即早期基因ICP27对单纯疱疹病毒1前mRNA剪接和多腺苷酸化的隐藏调控
作者：
Shuāng Táng;Amita Patel;P. Krause
与人类细胞相反，已知很少有HSV-1基因被剪接，尽管它们共享相同的前mRNA加工机制。在这里，通过对感染HSV-1和缺失感染性细胞培养蛋白27（ICP 27）（病毒复制所必需的两个病毒立即早期（IE）基因之一）的HSV-1突变病毒的细胞中的剪接点的全局分析，我们鉴定了数百个映射到先前已知的HSV-1剪接基因和先前未知的剪接基因的新的可变剪接点，其中大多数改变病毒基因的编码潜力。基于RNA-Seq和RT-PCR对这些新的可变剪接基因进行了定量和定性的剪接效率分析，结果表明，只有当ICP 27缺失时，这些新的剪接位点的剪接才是有效的;而在野生型HSV-1感染的细胞中，这些新的剪接点的剪接以基因/序列特异性的方式大部分被沉默，提示ICP 27不仅促进ICP 27靶向转录物的积累，而且通过抑制可变剪接确保功能编码序列的正确性。此外，ICP 27通过抑制新鉴定的内含子中的近端多聚腺苷酸化信号（PAS）的剪接和激活来切换主要病毒神经毒力因子ICP34.5的表达，揭示了病毒基因表达的新的调节机制。因此，通过病毒IE蛋白ICP 27，HSV-1选择剪接和多聚腺苷酸化机制以在裂解感染期间实现最佳病毒基因表达。另一方面，在ICP 27不存在时的潜伏感染期间，HSV-1可能利用宿主剪接机制来限制随机激活的抗原性病毒基因的表达，以实现免疫逃避。
In contrast to human cells, very few HSV-1 genes are known to be spliced, although the same pre-mRNA processing machinery is shared. Here, through global analysis of splice junctions in cells infected with HSV-1 and an HSV-1 mutant virus with deletion of infectious cell culture protein 27 (ICP27), one of two viral immediate early (IE) genes essential for viral replication, we identify hundreds of novel alternative splice junctions mapping to both previously known HSV-1 spliced genes and previously unknown spliced genes, the majority of which alter the coding potential of viral genes. Quantitative and qualitative splicing efficiency analysis of these novel alternatively spliced genes based on RNA-Seq and RT-PCR reveals that splicing at these novel splice sites is efficient only when ICP27 is absent; while in wildtype HSV-1 infected cells, the splicing of these novel splice junctions is largely silenced in a gene/sequence specific manner, suggesting that ICP27 not only promotes accumulation of ICP27 targeted transcripts but also ensures correctness of the functional coding sequences through inhibition of alternative splicing. Furthermore, ICP27 toggles expression of ICP34.5, the major viral neurovirulence factor, through inhibition of splicing and activation of a proximal polyadenylation signal (PAS) in the newly identified intron, revealing a novel regulatory mechanism for expression of a viral gene. Thus, through the viral IE protein ICP27, HSV-1 co-opts both splicing and polyadenylation machinery to achieve optimal viral gene expression during lytic infection. On the other hand, during latent infection when ICP27 is absent, HSV-1 likely takes advantages of host splicing machinery to restrict expression of randomly activated antigenic viral genes to achieve immune evasion.
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DOI：
10.1172/jci16126
发表时间：
2002-01
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
R. Whitley;B. Roizman
通讯作者：
R. Whitley;B. Roizman
影响因子：
16
作者：
Chiu AC;Suzuki HI;Wu X;Mahat DB;Kriz AJ;Sharp PA
通讯作者：
Sharp PA
影响因子：
10.5
作者：
Sandri-Goldin, RM
通讯作者：
Sandri-Goldin, RM
影响因子：
10.5
作者：
K. M. Wassarman;J. Steitz
通讯作者：
K. M. Wassarman;J. Steitz
影响因子：
3.7
作者：
Escudero-Paunetto L;Li L;Hernandez FP;Sandri-Goldin RM
通讯作者：
Sandri-Goldin RM