Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Proc Natl Acad Sci U S A. 2013 Feb 26;110(9):3519-24. doi: 10.1073/pnas.1300708110. Epub 2013 Feb 11.
Poxviruses are considered less dependent on host functions than other DNA viruses because of their cytoplasmic site of replication and large genomes, which encode enzymes for DNA and mRNA synthesis. Nevertheless, RNAi screens with two independent human genome-scale libraries have identified more than 500 candidate genes that significantly inhibited and a similar number that enhanced replication and spread of infectious vaccinia virus (VACV). Translational, ubiquitin-proteosome, and endoplasmic reticulum-to-Golgi transport functions, known to be important for VACV, were enriched in the siRNA-inhibiting group, and RNA polymerase II and associated functions were enriched in the siRNA-enhancing group. Additional findings, notably the inhibition of VACV spread by siRNAs to several nuclear pore genes, were unanticipated. Knockdown of nucleoporin 62 strongly inhibited viral morphogenesis, with only a modest effect on viral gene expression, recapitulating and providing insight into previous studies with enucleated cells.
痘病毒被认为比其他 DNA 病毒更少依赖于宿主功能，因为它们在细胞质中进行复制，并且基因组较大，可编码用于 DNA 和 mRNA 合成的酶。然而，使用两个独立的人类全基因组文库进行的 RNAi 筛选已经鉴定出 500 多个候选基因，这些基因显著抑制了传染性牛痘病毒 (VACV) 的复制和传播，而类似数量的基因则增强了其复制和传播。已知对 VACV 很重要的翻译、泛素-蛋白酶体和内质网到高尔基体运输功能在 siRNA 抑制组中富集，而 RNA 聚合酶 II 和相关功能在 siRNA 增强组中富集。其他发现，特别是核孔几个基因的 siRNA 抑制 VACV 传播，是出乎意料的。核孔蛋白 62 的敲低强烈抑制病毒形态发生，对病毒基因表达只有适度影响，这再现并为去核细胞的先前研究提供了深入了解。
