A Genome-Wide CRISPR-Cas9 Screen Identifies the Dolichol-Phosphate Mannose Synthase Complex as a Host Dependency Factor for Dengue Virus Infection.
Animals
CRISPR-Cas Systems
Chlorocebus aethiops
Dengue
/ virology
Dengue Virus
/ physiology
Endoplasmic Reticulum
/ metabolism
Fibroblasts
/ metabolism
Glycoproteins
/ metabolism
Glycosylation
Glycosylphosphatidylinositols
/ metabolism
HEK293 Cells
Humans
Mannose
/ chemistry
Mannosyltransferases
/ metabolism
Oligosaccharides
/ chemistry
RNA, Guide, Kinetoplastida
/ metabolism
RNA, Viral
/ chemistry
Vero Cells
Virus Replication
N-glycosylation
Zika virus
dengue fever
dengue virus
dolichol-phosphate mannose
glycoprotein folding
Journal
Journal of virology
ISSN: 1098-5514
Titre abrégé: J Virol
Pays: United States
ID NLM: 0113724
Informations de publication
Date de publication:
17 03 2020
17 03 2020
Historique:
received:
11
10
2019
accepted:
20
12
2019
pubmed:
10
1
2020
medline:
15
9
2020
entrez:
10
1
2020
Statut:
epublish
Résumé
Dengue virus (DENV) is a mosquito-borne flavivirus responsible for dengue disease, a major human health concern for which no specific therapies are available. Like other viruses, DENV relies heavily on the host cellular machinery for productive infection. In this study, we performed a genome-wide CRISPR-Cas9 screen using haploid HAP1 cells to identify host genes important for DENV infection. We identified DPM1 and -3, two subunits of the endoplasmic reticulum (ER) resident dolichol-phosphate mannose synthase (DPMS) complex, as host dependency factors for DENV and other related flaviviruses, such as Zika virus (ZIKV). The DPMS complex catalyzes the synthesis of dolichol-phosphate mannose (DPM), which serves as mannosyl donor in pathways leading to N-glycosylation, glycosylphosphatidylinositol (GPI) anchor biosynthesis, and C- or O-mannosylation of proteins in the ER lumen. Mutation in the DXD motif of DPM1, which is essential for its catalytic activity, abolished DPMS-mediated DENV infection. Similarly, genetic ablation of ALG3, a mannosyltransferase that transfers mannose to lipid-linked oligosaccharide (LLO), rendered cells poorly susceptible to DENV. We also established that in cells deficient for DPMS activity, viral RNA amplification is hampered and truncated oligosaccharides are transferred to the viral prM and E glycoproteins, affecting their proper folding. Overall, our study provides new insights into the host-dependent mechanisms of DENV infection and supports current therapeutic approaches using glycosylation inhibitors to treat DENV infection.
Identifiants
pubmed: 31915280
pii: JVI.01751-19
doi: 10.1128/JVI.01751-19
pmc: PMC7081898
pii:
doi:
Substances chimiques
Glycoproteins
0
Glycosylphosphatidylinositols
0
Oligosaccharides
0
RNA, Guide
0
RNA, Viral
0
Mannosyltransferases
EC 2.4.1.-
dolichyl-phosphate beta-D-mannosyltransferase
EC 2.4.1.83
Mannose
PHA4727WTP
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
Copyright © 2020 American Society for Microbiology.
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