A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry

• Published in: Viruses Vol. 13; no. 11; p. 2113
• Main Authors: Luu, Anh Phuong, Yao, Zhenlan, Ramachandran, Sangeetha, Azzopardi, Stephanie A, Miles, Linde A, Schneider, William M, Hoffmann, H-Heinrich, Bozzacco, Leonia, Garcia, Jr, Gustavo, Gong, Danyang, Damoiseaux, Robert, Tang, Hengli, Morizono, Kouki, Rudin, Charles M, Sun, Ren, Arumugaswami, Vaithilingaraja, Poirier, John T, MacDonald, Margaret R, Rice, Charles M, Li, Melody M H
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 20.10.2021; MDPI
• Subjects: A549 Cells; CRISPR; CRISPR activation; CRISPR-Cas Systems; Dengue fever; Fibroblasts; Genes; Genomes; GTP-Binding Proteins - genetics; GTP-Binding Proteins - metabolism; Guanosine triphosphatases; Host-pathogen interactions; Humans; Infections; Interferon; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Neural coding; p21-Activated Kinases - genetics; p21-Activated Kinases - metabolism; Proteins; proviral factors; Rho GTPases; rhoB GTP-Binding Protein - genetics; rhoB GTP-Binding Protein - metabolism; RhoV; Transcription; Transcriptional Coactivator with PDZ-Binding Motif Proteins - genetics; Transcriptional Coactivator with PDZ-Binding Motif Proteins - metabolism; Viral infections; Virions; Virus Internalization; Virus Replication; West Nile virus; WWTR1; Zika virus; Zika Virus - genetics; Zika Virus - physiology; Zika Virus Infection - enzymology; Zika Virus Infection - genetics; Zika Virus Infection - virology; WWTR1; CRISPR activation; Zika virus; Rho GTPases; RhoV; proviral factors
• ISSN: 1999-4915; 1999-4915
• 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.