Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095, USA.
Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA.
Viruses. 2021 Oct 20;13(11):2113. doi: 10.3390/v13112113.
Zika virus (ZIKV) is a re-emerging flavivirus that has caused large-scale epidemics. Infection during pregnancy can lead to neurologic developmental abnormalities in children. There is no approved vaccine or therapy for ZIKV. To uncover cellular pathways required for ZIKV that can be therapeutically targeted, we transcriptionally upregulated all known human coding genes with an engineered CRISPR-Cas9 activation complex in human fibroblasts deficient in interferon (IFN) signaling. We identified Ras homolog family member V () and WW domain-containing transcription regulator 1 () as proviral factors, and found them to play important roles during early ZIKV infection in A549 cells. We then focused on RhoV, a Rho GTPase with atypical terminal sequences and membrane association, and validated its proviral effects on ZIKV infection and virion production in SNB-19 cells. We found that RhoV promotes infection of some flaviviruses and acts at the step of viral entry. Furthermore, RhoV proviral effects depend on the complete GTPase cycle. By depleting Rho GTPases and related proteins, we identified RhoB and Pak1 as additional proviral factors. Taken together, these results highlight the positive role of RhoV in ZIKV infection and confirm CRISPR activation as a relevant method to identify novel host-pathogen interactions.
寨卡病毒（ZIKV）是一种重新出现的黄病毒，已导致大规模流行。孕妇感染寨卡病毒可导致儿童神经发育异常。目前尚无针对寨卡病毒的批准疫苗或疗法。为了发现可用于治疗寨卡病毒的细胞途径，我们在缺乏干扰素（IFN）信号的人成纤维细胞中用工程化的 CRISPR-Cas9 激活复合物转录上调了所有已知的人类编码基因。我们确定 Ras 同源家族成员 V（）和含有 WW 结构域的转录调节剂 1（）为前病毒因子，并发现它们在 A549 细胞中早期寨卡病毒感染中发挥重要作用。然后，我们专注于 RhoV，一种具有非典型末端序列和膜结合的 Rho GTPase，并在 SNB-19 细胞中验证了其对寨卡病毒感染和病毒粒子产生的前病毒效应。我们发现 RhoV 促进了一些黄病毒的感染，并作用于病毒进入的步骤。此外，RhoV 的前病毒效应取决于完整的 GTPase 循环。通过耗尽 Rho GTPases 和相关蛋白，我们确定 RhoB 和 Pak1 为另外的前病毒因子。总之，这些结果突出了 RhoV 在寨卡病毒感染中的积极作用，并证实了 CRISPR 激活是鉴定新的宿主-病原体相互作用的相关方法。
