A Global Interactome Map of the Dengue Virus NS1 Identifies Virus Restriction and Dependency Host Factors

Hafirassou, Mohamed Lamine; Meertens, Laurent; Umaña-Diaz, Claudia; Labeau, Athena; Dejarnac, Ophélie; Bonnet-Madin, Lucie; Kümmerer, Beate M.; Delaugerre, Constance; Roingeard, Philippe; Vidalain, Pierre‐Olivier; Amara, Ali

doi:10.1016/j.celrep.2018.01.038

Cited by 26 publications

(31 citation statements)

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“…Structural ZIKV proteins carry out the entry and membrane fusion steps of the viral life cycle 21 , while NS proteins cooperatively remodel ER membranes to form replication sites and synthesize viral RNA 22 . Despite their limited size and number, the functions of most of the NS proteins are poorly characterized 23 , as are their interactions with host lipids 24 and potentially hundreds of unique proteins 3,12,25 . While the enigmatic nature of the ZIKV NS proteins and their interactions presented challenges to defining a mechanistic basis for our lipidomics results, two lines of evidence led us to investigate NS4B as potentially important in altering lipid metabolism.…”

Section: Resultsmentioning

confidence: 99%

A global lipid map defines a network essential for Zika virus replication

Leier

Weinstein

Kyle

et al. 2020

Preprint

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26Zika virus (ZIKV), an arbovirus of global concern, remodels intracellular membranes to form 27 replication sites. How ZIKV dysregulates lipid networks to allow this, and consequences for 28 disease, is poorly understood. Here, we performed comprehensive lipidomics to create a lipid 29 network map during ZIKV infection. We found that ZIKV significantly alters host lipid 30 composition, with the most striking changes seen within subclasses of sphingolipids. Ectopic 31 expression of ZIKV NS4B protein resulted in similar changes, demonstrating a role for NS4B in 32 modulating sphingolipid pathways. Disruption of sphingolipid biosynthesis in various cell types, 33including human neural progenitor cells, blocked ZIKV infection. Additionally, the sphingolipid 34 ceramide redistributes to ZIKV replication sites and increasing ceramide levels by multiple 35 pathways sensitizes cells to ZIKV infection. Thus, we identify a sphingolipid metabolic network 36 with a critical role in ZIKV replication and show that ceramide flux is a key mediator of ZIKV 37 infection. Results 112 ZIKV alters the lipid landscape of host cells 113To understand how cellular lipid metabolism is altered by ZIKV infection, we carried out 114 a global lipidomic survey of the model Huh7 human hepatic carcinoma cell line 20 infected for 24 115 or 48 hours with Asian lineage ZIKV strain FSS13025 (Fig. 1a). Lipids extracted from 116 populations of mock and infected cells (n = 5 biological replicates per condition) were analyzed 117 with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) (Supplementary Data 118 1 and Supplementary Fig. 1a-f). We identified 340 lipid species spanning the phospholipid, 119 sphingolipid, glycerolipid, and sterol classes for which pairwise comparisons of normalized 120 abundance between mock and infected cells could be made (Supplementary Data 1). Of these, 121 80 species (23.5%) showed significant changes in abundance by 24 hours post-infection (hpi) 122 and 172 species (50.6%) were significantly altered by 48 hpi (P < 0.05, ANOVA or g-test) 123( Supplementary Data 1). Principal component analysis (PCA) of these observations confirmed 124 that infection status (mock or ZIKV) and timepoint (24 or 48 hpi) accounted most of these 125 changes, with changes in lipid composition between mock and infected cells increasing over 126 time (Fig. 1b). 127Next, we examined how ZIKV-induced changes in host lipid composition broke down by 128 subclass and species (Fig. 1c, d). A map of the pairwise correlations of all 340 species at 48 hpi 129 ( Supplementary Fig. 2a) revealed that lipid subclasses largely fell into two groups of species 130 that were either enriched or depleted in abundance ( Supplementary Fig. 2b), suggesting that 131 individual metabolic pathways are up-or down-regulated to create a specific lipid milieu around 132 the events of the viral replication cycle. Supporting this, many of the trends we observed were 133 consistent with earlier reports of functional roles for lipids during flavivirus infection. In ...

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Section: Resultsmentioning

confidence: 99%

A global lipid map defines a network essential for Zika virus replication

Leier

Weinstein

Kyle

et al. 2020

Preprint

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“…Notably, we find that under these conditions, capsid in the virion has aberrant electrophoretic mobility, suggestive of an altered conformation. In addition to Hsp70, other chaperones, such as Hsp90 or TRiC, may also facilitate virus protein folding and assembly (Geller et al, 2012; Hafirassou et al, 2018; Inoue et al, 2011). The precise dependence of a given virus family for distinct classes of chaperones may depend on their topology and folding pathway.…”

Section: Discussionmentioning

confidence: 99%

Zika Virus Dependence on Host Hsp70 Provides a Protective Strategy against Infection and Disease

et al. 2019

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SUMMARY The spread of mosquito-borne Zika virus (ZIKV), which causes neurological disorders and micro-cephaly, highlights the need for countermeasures against sudden viral epidemics. Here, we tested the concept that drugs targeting host proteostasis pro-vide effective antivirals. We show that different cyto-solic Hsp70 isoforms are recruited to ZIKV-induced compartments and are required for virus replication at pre- and post-entry steps. Drugs targeting Hsp70 significantly reduce replication of different ZIKV strains in human and mosquito cells, including hu-man neural stem cells and a placental trophoblast cell line, at doses without appreciable toxicity to the host cell. By targeting several ZIKV functions, including entry, establishment of active replication complexes, and capsid assembly, Hsp70 inhibitors are refractory to the emergence of drug-resistant virus. Importantly, these drugs protected mouse models from ZIKV infection, reducing viremia, mor-tality, and disease symptoms. Hsp70 inhibitors are thus attractive candidates for ZIKV therapeutics with the added benefit of a broad spectrum of action.

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“…For vertebrate-and insect-infecting viruses, RACK1 is known to be targeted during replication. For example, dengue virus NS1 protein interacts with RACK1 to facilitate virus replication without affecting viral translation (Hafirassou et al, 2017), although the underlying mechanism of RACK1 in dengue virus replication remains unknown. The interaction of infectious bursal disease virus protein VP5 with RACK1 inhibits apoptosis and thereby keeps cells suitable for virus replication (Lin et al, 2015).…”

Section: Researchmentioning

confidence: 99%

Hijacking a host scaffold protein, RACK1, for replication of a plant RNA virus

2018

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Receptor for activated C kinase 1 (RACK1) is strictly conserved across eukaryotes and acts as a versatile scaffold protein involved in various signaling pathways. Plant RACK1 is known to exert important functions in innate immunity against fungal and bacterial pathogens. However, the role of the RACK1 in plant-virus interactions remains unknown. Here, we addressed the role of RACK1 of Nicotiana benthamiana during infection by red clover necrotic mosaic virus (RCNMV), a plant positive-stranded RNA virus. NbRACK1 was shown to be recruited by the p27 viral replication protein into endoplasmic reticulum-derived aggregated structures (possible replication sites). Downregulation of NbRACK1 by virus-induced gene silencing inhibited viral cap-independent translation and p27-mediated reactive oxygen species (ROS) accumulation, which are prerequisite for RCNMV replication. We also found that NbRACK1 interacted with a host calcium-dependent protein kinase (NbCDPKiso2) that activated a ROS-generating enzyme. Interestingly, NbRACK1 was required for the interaction of p27 with NbCDPKiso2, suggesting that NbRACK1 acts as a bridge between the p27 viral replication protein and NbCDPKiso2. Collectively, our findings provide an example of a viral strategy in which a host multifaceted scaffold protein RACK1 is highjacked for promoting viral protein-triggered ROS production necessary for robust viral replication.

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A Global Interactome Map of the Dengue Virus NS1 Identifies Virus Restriction and Dependency Host Factors

Abstract: In the originally published version of this article, Figure S1 in the Supplemental Information became unreadable during the production process. Figure S1 has now been corrected online. The Cell Reports team regrets this error.

Cited by 26 publications

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A global lipid map defines a network essential for Zika virus replication

A global lipid map defines a network essential for Zika virus replication

Zika Virus Dependence on Host Hsp70 Provides a Protective Strategy against Infection and Disease

Hijacking a host scaffold protein, RACK1, for replication of a plant RNA virus

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