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Cited by 411Mosquito-borne diseases and controlViral Infectious Diseases and Gene Expression in InsectsViral Infections and Vectors

Genetic dissection of Flaviviridae host factors through genome-scale CRISPR screens

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Marceau, Caleb D., Puschnik, Andreas S., Majzoub, Karim, Ooi, Yaw Shin, Brewer, Susan M., Fuchs, Gabriele, Swaminathan, Kavya, Mata, Miguel A. +3 more·2016·Nature
DOI 10.1038/nature18631PMC PMC4964798
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Abstract

The Flaviviridae are a family of viruses that cause severe human diseases. For example, dengue virus (DENV) is a rapidly emerging pathogen causing an estimated 100 million symptomatic infections annually worldwide. No approved antivirals are available to date and clinical trials with a tetravalent dengue vaccine showed disappointingly low protection rates. Hepatitis C virus (HCV) also remains a major medical problem, with 160 million chronically infected patients worldwide and only expensive treatments available. Despite distinct differences in their pathogenesis and modes of transmission, the two viruses share common replication strategies. A detailed understanding of the host functions that determine viral infection is lacking. Here we use a pooled CRISPR genetic screening strategy to comprehensively dissect host factors required for these two highly important Flaviviridae members. For DENV, we identified endoplasmic-reticulum (ER)-associated multi-protein complexes involved in signal sequence recognition, N-linked glycosylation and ER-associated degradation. DENV replication was nearly completely abrogated in cells deficient in the oligosaccharyltransferase (OST) complex. Mechanistic studies pinpointed viral RNA replication and not entry or translation as the crucial step requiring the OST complex. Moreover, we show that viral non-structural proteins bind to the OST complex. The identified ER-associated protein complexes were also important for infection by other mosquito-borne flaviviruses including Zika virus, an emerging pathogen causing severe birth defects. By contrast, the most significant genes identified in the HCV screen were distinct and included viral receptors, RNA-binding proteins and enzymes involved in metabolism. We found an unexpected link between intracellular flavin adenine dinucleotide (FAD) levels and HCV replication. This study shows notable divergence in host-depenency factors between DENV and HCV, and illuminates new host targets for antiviral therapy.

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  • Pooled CRISPR screening is a powerful and widely used method for identifying genes involved in biological mechanisms such as cell proliferation, drug resistance, and viral infection.

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Authors
Marceau, C. D., Puschnik, A. S., Majzoub, K., Ooi, Y. S., Brewer, S. M., Fuchs, G., Swaminathan, K., Mata, M. A., Elias, J. E., Sarnow, P., & Carette, J. E.
Affiliations
Stanford University (US)
Corresponding author
Carette, Jan E.
ORCID
Venue
Nature
Publisher
Springer Science and Business Media LLC
Published
2016-06-17
Volume
535
Issue
7610
Pages
159-163
ISSN
  • 0028-0836 (ISSN-L)
  • 1476-4687
DOI
10.1038/nature18631
PMC
PMC4964798
OpenAlex
W2439020788
Cited by
411
References
2
Citations by year
2026: 20 · 2025: 32 · 2024: 35 · 2023: 32 · 2022: 29 · 2021: 48 · 2020: 56 · 2019: 46 · 2018: 60 · 2017: 38 · 2016: 15
FWCI
45.24
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
Funders
  • National Science Foundation
  • Boehringer Ingelheim Fonds
  • National Institutes of Health (5u19ai109662-05, AI109662)
  • National Institute of Allergy and Infectious Diseases (AI109662, DP2AI104557, U19-AI109662, T32AI007328)
Topics
  • Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Mosquito-borne diseases and control
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
  • Health Sciences › Medicine › Infectious Diseases › Viral Infections and Vectors
Keywords
dengue virus, hepatitis C virus, Flaviviridae, host factors, oligosaccharyltransferase, signal peptidase complex, N-glycosylation, viral RNA replication, viral nonstructural proteins, FAD biosynthesis, RNA-binding proteins, viral entry, mosquito-borne flaviviruses, West Nile virus, Zika virus, CRISPR screen
MeSH terms
Zika Virus, Dengue Virus, Endoplasmic Reticulum, Flavin-Adenine Dinucleotide, Glycosylation, Hexosyltransferases, Humans, Membrane Proteins, Protein Binding, Receptors, Virus, Virus Replication, Genome, Human, Hepacivirus, RNA-Binding Proteins, Viral Nonstructural Proteins, Flavivirus Infections, Protein Sorting Signals, Host-Pathogen Interactions, Drug Discovery, Host-Derived Cellular Factors, Molecular Targeted Therapy, Endoplasmic Reticulum-Associated Degradation, CRISPR-Cas Systems

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