Secreted dengue virus NS1 from infection is predominantly dimeric and in complex with high-density lipoprotein

Liang, Alvin Chew Bing; Ngoh, An Qi; Phoo, Wint Wint; Chan, Wing Ki; Ser, Zheng; Lim, Shiao See; Weng, Meng Jie Grace; Watanabe, Sumio; Choy, Milly M.; Low, Guek Hong Jenny; Ooi, Eng Eong; Sobota, Radoslaw M.; Vasudevan, Subhash G.; Luo, Dahai

doi:10.1101/2022.04.06.487425

Cited by 5 publications

(19 citation statements)

References 66 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Compared to the available structures, we present a new subset of antibody-NS1 complex structures at antigen-binding sites between the wing and β-ladder of the NS1 protein. Lastly, we showed that ZIKV isNS1 is found in complex with apoA1; they are dimers docked onto HDL, similar to earlier reports for DENV 30,33 .…”

Section: Introductionsupporting

confidence: 90%

Structural basis of Zika virus NS1 multimerization and human antibody recognition

Luo,

Chew,

Ngoh

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

The Zika virus (ZIKV) is a mosquito-transmitted flavivirus related to the dengue virus that has emerged in recent global outbreaks, with previously unreported neurological or birth abnormalities. There remains a large, unmet need for efficacious vaccines and antiviral agents against ZIKV. Flavivirus non-structural protein 1 (NS1) is the only secreted viral protein that has proven useful in diagnostics. However, its physiologically relevant structural forms and pathogenesis mechanisms remain unclear. While many antibodies targeting NS1 with mapped immunodomain epitopes have been reported, antibody-NS1 structures are starting to emerge, which may guide vaccine and therapeutic design. Here, we present high-resolution cryoEM structures of ZIKV recombinant secreted NS1 (rsNS1) and its complexes with three human monoclonal antibodies, as well as evidence of ZIKV infection-derived secreted NS1 (isNS1) binding to HDL. We showed that ZIKV rsNS1 forms tetramers and filamentous repeats of tetramers. The studied antibodies (AA12, EB9, GB5) did not disrupt the ZIKV NS1 tetramers as they bound to the wing and connector subdomain to the β-ladder. Our study reveals new insights into NS1 multimerization, highlights the need to distinguish the polymorphic nature of rsNS1 and isNS1, and expands the mechanistic basis of the protection conferred by antibodies targeting NS1.

show abstract

Section: Introductionsupporting

confidence: 90%

Structural basis of Zika virus NS1 multimerization and human antibody recognition

Luo,

Chew,

Ngoh

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…As there were marked differences between ApoA1 and cholesterol subfractions between patients with DF and DHF, and since it was shown that ApoA1 binds to DENV NS1 [24], we wished to evaluate the possible effects of ApoA1 and NS1 together on primary human . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.…”

Section: Effect Of Apoa1 and Ns1 On Primary Human Monocytes In Induci...mentioning

confidence: 99%

“…As there were marked differences between ApoA1 and cholesterol subfractions between patients with DF and DHF, and since it was shown that ApoA1 binds to DENV NS1 [24], we wished to evaluate the possible effects of ApoA1 and NS1 together on primary human monocytes. In order to determine the effect of ApoA1 along with NS1, primary monocytes of 6 individuals were co-cultured with varying concentrations of natural human ApoA1 (specified by the manufacturer as endotoxin free) at various concentrations (195μg/ml, 100μg/ml, 65μg/ml, 10μg/ml, and 1.68μg/ml) together with DENV1 NS1 antigen, that was specified by the manufacturer as endotoxin free (1μg/ml) (Native antigen, UK) for 16 hours.…”

Section: Effect Of Apoa1 and Ns1 On Primary Human Monocytes In Induci...mentioning

confidence: 99%

“…It was recently shown that the lipid composition of NS1 hexamer was different in NS1 antigens associated with increased virulence, compared to those who were associated with milder disease [20]. NS1 complex in the supernatants of AD293 transfected cell lines, was shown to coimmunoprecipitate with apolipoprotein 1 (ApoA1) [22] and NS1 antigen was shown to directly interact with ApoA1 in patients [23,24]. Interaction of ApoA1 opposes the action of NS1 on lipid rafts in cells and thereby reducing the infectivity of the DENV [23].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Apolipoprotein A1 levels and its association with NS1 in the pathogenesis of acute dengue

Jayathilaka

Gomes

Jeewandara

et al. 2022

Preprint

View full text Add to dashboard Cite

Background The dengue NS1 antigen is a secretory protein and was shown to associate with apolipoprotein A1 (ApoA1). Therefore, we sought to investigate the changes in ApoA1 levels in patients with varying severity of acute dengue and then proceeded to investigate how ApoA1 and NS1 interactions affect cytokine production from primary human monocytes. Methodology/Principal Findings Serial measurements of ApoA1, viral loads, NS1 antigen levels and lipid profiles were done in adult patients with dengue fever (DF= 21) and dengue haemorrhagic fever (DHF =28). To investigate the effect of ApoA1-NS1 interactions in cytokine production from primary human monocytes, cells were harvested from healthy individuals (n=6) and co-cultured with varying concentrations of ApoA1 and levels of IL-6 and TNF-α measured in culture supernatants. The ApoA1 levels were significantly lower in patients with DHF on day 5 of illness (p=0.04) compared to those with DF. ApoA1 levels did not show any correlation with either the NS1 antigen levels or the viral loads in patients with DF or DHF, although they did significantly and inversely correlate with liver transaminases, AST (Spearmans R=-0.55, p <0.0001) and ALT (Spearmans R=-0.49, p<0.0001) in patients with DF. However, a significant correlation was not seen between ApoA1 levels and AST levels (Spearmans R=-0.45, p=0.09) in patients with DHF and ALT levels (Spearman’s R=0.0.5, p=0.64). HDL and LDL levels were significantly lower on day 5 of the illness in patients with DHF compared to those with DF. Co-culture of NS1 and ApoA1 resulted in an increased IL-6 production (but not TNFα) in culture supernatants in a dose dependent manner. Conclusions/Significance HDL, LDL and ApoA1 levels were significantly lower in those who have severe dengue, especially in the critical phase and ApoA1 levels inversely correlated with the extent of rise in liver enzymes. While co-culture of ApoA1 with NS1 in primary human monocytes induced high IL-6 levels in a dose dependent manner, this was not seen for TNFα suggesting that the interaction of ApoA1 with NS1 could give rise to different outcomes.

show abstract

“…Similarly, ZIKV infection modulates the placenta lipidome for intracellular membrane reorganization (like formation of vesicle pockets on the ER luminal side visualized by TEM imaging) and viral replication . Several reports have also suggested that NS1 affects cholesterol metabolism and triggers proinflammatory responses by interacting with HDL (high density lipoprotein, responsible for reverse cholesterol transport). − Recent cryo-EM studies have highlighted the formation of higher order oligomeric states (dimers and hexameric) of NS1 with HDL. , All these studies emphasize the significance of NS1 interaction with lipids and lipoproteins for replication and disease progression inside the host cells. Hence, a molecular level exploration of the interaction of NS1 with cholesterol containing membrane is necessary to understand the formation of NS1-HDL complexes.…”

Section: Introductionmentioning

confidence: 99%

Differences in the Membrane-Binding Properties of Flaviviral Nonstructural 1 (NS1) Protein: Comparative Simulations of Zika and Dengue Virus NS1 Proteins in Explicit Bilayers

Muthukumaran,

Sankararamakrishnan

2024

ACS Bio Med Chem Au

View full text Add to dashboard Cite

NS1 in flaviviruses is the only nonstructural protein that is secretory and interacts with different cellular components of the host cell membrane. NS1 is localized in the ER as a dimer to facilitate viral replication. Crystal structures of NS1 homologues from zika (ZIKV) and dengue (DENV) viruses have revealed the organization of different domains in NS1 dimers. The β-roll and the connector and intertwined loop regions of wing domains of NS1 have been shown to interact with the membranes. In this study, we have performed multiple molecular dynamics (MD) simulations of ZIKV and DENV NS1 systems in apo and in POPE bilayers with different cholesterol concentrations (0, 20 and 40%). The NS1 protein was placed just above the membrane surface, and for each NS1-membrane system two to three independent simulations with 600 ns production run were performed. At the end of the production runs, ZIKV NS1 inserts deeper inside the membrane compared to the DENV counterpart. Unlike ZIKV NS1, the orientation of DENV NS1 is asymmetric in which one of the chains in the dimer interacts with the membrane while the other is more exposed to the solvent. The β-roll region in ZIKV NS1 penetrates beyond the headgroup region and interacts with the lipid acyl chains while the C-terminal region barely interacts with the headgroup. Specific residues in the intertwined region deeply penetrate inside the membrane. The role of charged and aromatic residues of ZIKV NS1 in strongly interacting with the membrane components is revealed. The presence of cholesterol affects the extent of insertion in the membrane and interaction of individual residues. Overall, membrane-binding properties of ZIKV NS1 significantly differ from its counterpart in DENV. The differences found in the binding and insertion of NS1 can be used to design drugs and novel antibodies that can be flavivirus specific.

show abstract

Secreted dengue virus NS1 from infection is predominantly dimeric and in complex with high-density lipoprotein

Cited by 5 publications

References 66 publications

Structural basis of Zika virus NS1 multimerization and human antibody recognition

Structural basis of Zika virus NS1 multimerization and human antibody recognition

Apolipoprotein A1 levels and its association with NS1 in the pathogenesis of acute dengue

Differences in the Membrane-Binding Properties of Flaviviral Nonstructural 1 (NS1) Protein: Comparative Simulations of Zika and Dengue Virus NS1 Proteins in Explicit Bilayers

Contact Info

Product

Resources

About