A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry
Anh Phuong Luu,
Zhenlan Yao,
Sangeetha Ramachandran,
Stephanie A. Azzopardi,
Linde A. Miles,
William M. Schneider,
H.-Heinrich Hoffmann,
Leonia Bozzacco,
Gustavo Garcia,
Danyang Gong,
Robert Damoiseaux,
Hengli Tang,
Kouki Morizono,
Charles M. Rudin,
Ren Sun,
Vaithilingaraja Arumugaswami,
John T. Poirier,
Margaret R. MacDonald,
Charles M. Rice,
Melody M. H. Li
Affiliations
Anh Phuong Luu
Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095, USA
Zhenlan Yao
Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095, USA
Sangeetha Ramachandran
Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095, USA
Stephanie A. Azzopardi
Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA
Linde A. Miles
Human Oncology & Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA
William M. Schneider
Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA
H.-Heinrich Hoffmann
Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA
Leonia Bozzacco
Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA
Gustavo Garcia
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90095, USA
Danyang Gong
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90095, USA
Robert Damoiseaux
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90095, USA
Hengli Tang
Department of Biological Science, Florida State University, Tallahassee, FL 32306, USA
Kouki Morizono
Division of Hematology and Oncology, Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA
Charles M. Rudin
Druckenmiller Center for Lung Cancer Research and Department of Medicine, Thoracic Oncology Service, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA
Ren Sun
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90095, USA
Vaithilingaraja Arumugaswami
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90095, USA
John T. Poirier
Laura and Isaac Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016, USA
Margaret R. MacDonald
Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA
Charles M. Rice
Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA
Melody M. H. Li
Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095, USA
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 (RhoV) and WW domain-containing transcription regulator 1 (WWTR1) 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.
