Potential of Peptide-Based Non-Structural Protein 1 (NS1) Inhibitor in Obstructing Dengue Virus (DENV) Replication

Introduction: Dengue Virus (DENV) is the pathogen for human dengue fever and is responsible for 390 million infections per year. The viral genome produces about 10 viral protein products, one of them being NS1. The NS1 protein plays a key role in viral replication and stimulation of humoral immune cells, thus being the perfect candidate to create an effective antiviral drug or vaccine for dengue Methods: Dengue Virus (DENV) is the pathogen for human dengue fever and is responsible for 390 million infections per year. The viral genome produces about 10 viral protein products, one of them being NS1. The NS1 protein plays a key role in viral replication and stimulation of humoral immune cells, thus being the perfect candidate to create an effective antiviral drug or vaccine for dengue Conclusion: The review established promising results of using peptide-based intervention on NS1. Further in vivo and randomized controlled trials are advised to solidify the applicability and biosafety of the intervention

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Diterpenes/Diterpenoids and Their Derivatives as Potential Bioactive Leads against Dengue Virus: A Computational and Network Pharmacology Study

Molecules ◽

10.3390/molecules26226821 ◽

2021 ◽

Vol 26 (22) ◽

pp. 6821

Author(s):

Rasel Ahmed Khan ◽

Rajib Hossain ◽

Abolghasem Siyadatpanah ◽

Khattab Al-Khafaji ◽

Abul Bashar Ripon Khalipha ◽

...

Keyword(s):

Dengue Virus ◽

Dengue Fever ◽

Computational Study ◽

Viral Proteins ◽

Network Pharmacology ◽

Binding Affinities ◽

Ns1 Protein ◽

Structural Protein ◽

E Protein

Dengue fever is a dangerous infectious endemic disease that affects over 100 nations worldwide, from Africa to the Western Pacific, and is caused by the dengue virus, which is transmitted to humans by an insect bite of Aedes aegypti. Millions of citizens have died as a result of dengue fever and dengue hemorrhagic fever across the globe. Envelope (E), serine protease (NS3), RNA-directed RNA polymerase (NS5), and non-structural protein 1 (NS1) are mostly required for cell proliferation and survival. Some of the diterpenoids and their derivatives produced by nature possess anti-dengue viral properties. The goal of the computational study was to scrutinize the effectiveness of diterpenoids and their derivatives against dengue viral proteins through in silico study. Methods: molecular docking was performed to analyze the binding affinity of compounds against four viral proteins: the envelope (E) protein, the NS1 protein, the NS3 protein, and the NS5 protein. Results: among the selected drug candidates, triptolide, stevioside, alepterolic acid, sphaeropsidin A, methyl dodovisate A, andrographolide, caesalacetal, and pyrimethamine have demonstrated moderate to good binding affinities (−8.0 to −9.4 kcal/mol) toward the selected proteins: E protein, NS3, NS5, and NS1 whereas pyrimethamine exerts −7.5, −6.3, −7.8, and −6.6 kcal/mol with viral proteins, respectively. Interestingly, the binding affinities of these lead compounds were better than those of an FDA-approved anti-viral medication (pyrimethamine), which is underused in dengue fever. Conclusion: we can conclude that diterpenoids can be considered as a possible anti-dengue medication option. However, in vivo investigation is recommended to back up the conclusions of this study.

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Critical role of Dengue Virus NS1 protein in viral replication

Virologica Sinica ◽

10.1007/s12250-014-3459-1 ◽

2014 ◽

Vol 29 (3) ◽

pp. 162-169 ◽

Cited By ~ 15

Author(s):

Jingjing Fan ◽

Yi Liu ◽

Zhiming Yuan

Keyword(s):

Dengue Virus ◽

Viral Replication ◽

Critical Role ◽

Ns1 Protein

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Determinants of Dengue Virus NS4A Protein Oligomerization

Journal of Virology ◽

10.1128/jvi.00546-15 ◽

2015 ◽

Vol 89 (12) ◽

pp. 6171-6183 ◽

Cited By ~ 22

Author(s):

Chia Min Lee ◽

Xuping Xie ◽

Jing Zou ◽

Shi-Hua Li ◽

Michelle Yue Qi Lee ◽

...

Keyword(s):

Amino Acids ◽

Dengue Virus ◽

Viral Replication ◽

Transmembrane Domain ◽

Antiviral Drug ◽

Protein Oligomerization ◽

Dependent Manner ◽

Wild Type ◽

Replication Complex ◽

Host Membrane

ABSTRACTFlavivirus NS4A protein induces host membrane rearrangement and functions as a replication complex component. The molecular details of how flavivirus NS4A exerts these functions remain elusive. Here, we used dengue virus (DENV) as a model to characterize and demonstrate the biological relevance of flavivirus NS4A oligomerization. DENV type 2 (DENV-2) NS4A protein forms oligomers in infected cells or when expressed alone. Deletion mutagenesis mapped amino acids 50 to 76 (spanning the first transmembrane domain [TMD1]) of NS4A as the major determinant for oligomerization, while the N-terminal 50 residues contribute only slightly to the oligomerization. Nuclear magnetic resonance (NMR) analysis of NS4A amino acids 17 to 80 suggests that residues L31, L52, E53, G66, and G67 could participate in oligomerization. Ala substitution for 15 flavivirus conserved NS4A residues revealed that these amino acids are important for viral replication. Among the 15 mutated NS4A residues, 2 amino acids (E50A and G67A) are located within TMD1. Both E50A and G67A attenuated viral replication, decreased NS4A oligomerization, and reduced NS4A protein stability. In contrast, NS4A oligomerization was not affected by the replication-defective mutations (R12A, P49A, and K80A) located outside TMD1.transcomplementation experiments showed that expression of wild-type NS4A alone was not sufficient to rescue the replication-lethal NS4A mutants. However, the presence of DENV-2 replicons could partially restore the replication defect of some lethal NS4A mutants (L26A and K80A), but not others (L60A and E122A), suggesting an unidentified mechanism governing the outcome of complementation in a mutant-dependent manner. Collectively, the results have demonstrated the importance of TMD1-mediated NS4A oligomerization in flavivirus replication.IMPORTANCEWe report that DENV NS4A forms oligomers. Such NS4A oligomerization is mediated mainly through amino acids 50 to 76 (spanning the first transmembrane domain [TMD1]). The biological importance of NS4A oligomerization is demonstrated by results showing that mutations of flavivirus conserved residues (E50A and G67A located within TMD1) reduced the oligomerization and stability of the NS4A protein, leading to attenuated viral replication. A systematic mutagenesis analysis demonstrated that flavivirus conserved NS4A residues are important for DENV replication. A successfultranscomplementation of replication-lethal NS4A mutant virus requires wild-type NS4A in the context of the viral replication complex. The wild-type NS4A protein alone is not sufficient to rescue the replication defect of NS4A mutants. Intriguingly, distinct NS4A mutants yielded different complementation outcomes in the replicon-containing cells. Overall, the study has enhanced our understanding of flavivirus NS4A at the molecular level. The results also suggest that inhibitor blocking of NS4A oligomerization could be explored for antiviral drug discovery.

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Immunogenicity of Recombinant DNA Vaccine Encoding Non-Structural Protein-1 Dengue Virus Serotype-2 in Balb/c Mice

Microbiology Indonesia ◽

10.5454/mi.15.2.4 ◽

2021 ◽

Vol 15 (2) ◽

pp. 4

Author(s):

Elitha Pulungan

Keyword(s):

Immune Response ◽

Dengue Virus ◽

Dna Vaccine ◽

Humoral Immune Response ◽

Structural Proteins ◽

Ns1 Protein ◽

Structural Protein ◽

Humoral Immune ◽

Virus Serotype ◽

Positive Strand Rna

Background: Dengue Hemorrhagic Fever (DHF) is an infectious disease caused by the dengue virus (DENV) which spread widely in tropical and subtropical regions of the world. DENV is a single-positive strand RNA virus with a genome size of ± 11kb which encodes three structural proteins, seven non-structural proteins, and two untranslated regions (UTR). The non-structural protein-1 (NS1) of DENV is known to have important role in dengue pathogenesis also promising to be developed as dengue vaccine. Lately, novel vaccine approach by DNA immunization have given new perspective for a safe, stable, and immunogenic vaccine platform. Previously, we have successfully construct DNA vaccine encoding NS1 protein of DENV2 (pUNS1) which express recombinant NS1 protein in-vitro. Thus, in this current study the ability of pUNS1 to induce humoral immune response will be further analyzed by in mice immunization. Methods: Sixteen BALB/c mice aged of 4 weeks were immunized 3 times with 100 µg of pUNS1 or pUMVC4a on 2 week time interval. Blood sampling was carried out just before immunization and termination was done 2 week after last immunization. Titer from individual mice sera against DENV-2 were measure with in-house ELISA. Results: IgG against NS1 protein of DENV2 titer from mice group immunized with recombinant pUNS1 shown high ELISA absorbancies, 5 times higher than pUMVC4a group. This result suggest the ability of pUNS1 to induce humoral immune response against NS1 DENV-2 in-vivo. Conclusion: Recombinant pUNS1 can induce humoral immune response in mice.

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Blood glucose promotes dengue virus infection in the mosquito Aedes aegypti

Parasites & Vectors ◽

10.1186/s13071-021-04877-1 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Shih-Che Weng ◽

Po-Nien Tsao ◽

Shin-Hong Shiao

Keyword(s):

Blood Glucose ◽

Aedes Aegypti ◽

Dengue Virus ◽

Viral Replication ◽

Dengue Fever ◽

Virus Transmission ◽

Hemorrhagic Fever ◽

Co Morbidity ◽

The Impact ◽

Blood Meals

Abstract Background Dengue fever is the most rapidly spreading mosquito-borne viral disease globally. More than 2.5 billion people live in dengue-endemic areas. Previous studies suggested an interrelationship between diabetes mellitus (DM) and dengue hemorrhagic fever (DHF). Conversely, glycolysis is a critical metabolic pathway for optimal dengue virus (DENV) replication. However, little is known concerning the effect of glucose on DENV replication in mosquitoes. In this study, we investigated the impact of glucose on DENV replication in mosquitoes Aedes aegypti. Methods Mosquitoes (Ae. aegypti UGAL/Rockefeller strain) were orally infected with DENV (serotype 2, 16681 strain) through infectious blood feeding. The DENV infection and transmission rates were determined by examining mosquito bodies and saliva, respectively, for DENV positivity at different time points after infection. In addition, a reverse genetic approach was applied by introducing double-stranded RNA against genes of interest into the mosquitoes to inhibit gene expression. Results Our data revealed a significant increase of DENV genome levels in mosquitoes consuming an infectious blood meal supplemented with glucose, suggesting that blood glucose is an important factor for viral replication. Interestingly, a significant increase of DENV E protein levels was detected in the saliva 4 days faster in mosquitoes that consumed infectious blood meals supplemented with glucose than in those consuming infectious blood meals alone. Furthermore, we perform RNAi to silence AKT or TOR and investigate the molecular mechanism regulating the glucose-mediated enhancement of viral replication. Silencing of AKT or TOR significantly reduced DENV titers in mosquitoes. Conclusions This study suggested that blood glucose is beneficial to DENV replication and that it facilitates virus transmission in mosquitoes via AKT and TOR signaling. Therefore, our results strengthen our understanding of dengue fever and DM co-morbidity and possibly reveal new targets for specific antiviral therapies. Graphical abstract

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OVERVIEW OF NUCLEAR FACTOR-KB (NF-KB) AND NON-STRUCTURAL PROTEIN 1 (NS1) IN PATIENTS WITH DENGUE FEVER IN PREMIER HOSPITAL, SURABAYA

Indonesian Journal of Tropical and Infectious Disease ◽

10.20473/ijtid.v7i5.9955 ◽

2019 ◽

Vol 7 (5) ◽

pp. 109

Author(s):

Ni Nyoman Budiutari ◽

Yoes Prijatna Dachlan ◽

Jusak Nugraha

Keyword(s):

Transcription Factors ◽

Viral Replication ◽

Dengue Fever ◽

Interleukin 1 ◽

Dengue Infection ◽

Sandwich Elisa ◽

Rapid Test ◽

Host Cells ◽

Ns1 Protein ◽

Structural Protein

Dengue fever (DF) is an acute viral fever caused by RNA virus that is transmitted by Aedes aegypti and Aedes albopictus mosquitoes. DF is also called viral arthropod-borne disease and is accompanied by headaches, joint and muscle pain. The main target of dengue infection is macrophages or monocytes and dendritic cells (DC). Infected DC is caused the viral replication and the endocytosis into endosomal, easier, thus inducing the activation of NF-ĸB transcription factor to produce proinflammatory cytokines such as Tumor Necrosis Factor-α (TNF-α), Interleukin-1 (IL-1), IL-6, IL-12 and chemokine. NF-kB is one of the transcription factors involved in the regulation of the expression of various cytokines, chemokines and anti/pro-apoptotic proteins during infection and act as indicator of disease severity. Infected DC cells are secreted NS1 protein which is the co-factor needed for viral replication and can be detected in the first eight days. The level will be higher in the initial phase of fever. The purpose of this study was to analyze the description of NF-kB and NS1 levels in the serum of patients with dengue fever through observational analytic studies through a cross-sectional approach. This study was done on 40 patients with dengue fever and 10 healthies people as negative controls. NS1 was analyzed in serum of Panbio rapid test and NF-kB level were measured by sandwich ELISA. The results are showed positive and negative NS1 results in dengue fever patients. The average NF-kB serum level in dengue fever patients was found to be higher than the control. NF-ĸB level in negative NS1 was higher than the NS1 positive group. It is showed that NS1 is detected both in the acute phase. The detection of NF-ĸB is showed the involvement of transcription factors in the development of dengue virus infection and has a protective role for host cells.

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In vitro and in silico Anti-dengue activity of Supercritical extract of medicinal plants against Dengue serotype-2

Research Journal of Pharmacy and Technology ◽

10.52711/0974-360x.2021.01025 ◽

2021 ◽

pp. 5895-5902

Author(s):

Sulochana Kaushik ◽

Lalit Dar ◽

Samander Kaushik ◽

Ramesh Kumar ◽

Devender Kumar ◽

...

Keyword(s):

Dengue Virus ◽

In Silico ◽

Antiviral Drug ◽

Docking Study ◽

Tinospora Cordifolia ◽

Ns1 Protein ◽

Toxic Dose ◽

Simplex Virus ◽

Therapeutic Compounds

Dengue is transmitted by female Aedes mosquitoes. It has been reported that about 2.5 billion peoples are at the risk of dengue virus. Millions of cases of dengue virus occur worldwide each year. There is no antiviral drug available still. Hence, the researchers are in the search of new anti-dengue drugs from natural products. The present study is aimed to determine the anti-dengue activity of supercritical extracts of Andrographis paniculata, Berberis vulgaris, Carica papaya, Euphorbia hirta, Phyllanthus niruri and Tinospora cordifolia in vitro and in silico. The cell viability was evaluated in C6/36 cells line by using MTT assay using a microplate reader at 595 nm. The maximum non-toxic dose of C. papaya and B. vulgaris extracts were reported as 46.87µg/ml, 31.25µg/ml for A. paniculata, P. niruri and E. hirta and 23.43 µg/ml, for T. cordifolia. Further, the anti-dengue activity of plants extract was analyzed by real-time RT-PCR. 100 copies/ml of DENV-2 virus was used for the antiviral assay. A. paniculata supercritical extract showed complete inhibition against the dengue-2 virus at 40ºC temperature and 15Mpa pressure. The other plant extracts showed the inhibition in order of T. cordifolia (83.44%) > C. papaya (34.71%) > E. hirta (28.70%) whereas P. niruri and B. vulgaris failed to inhibit dengue virus. Andrographolide a major compound present in A. paniculata has been reported to have antiviral activity against hepatitis B, C virus, herpes simplex virus, influenza virus, chikungunya virus, dengue virus 2 and 4. Results of molecular docking showed that the interaction between andrographolide and NS1protein shows the maximum binding energy -7.30 Kcal/mol. The docking study was conducted to validate the result against the anti-dengue activity using dengue NS1 protein with andrographolide. It was concluded that A. paniculata could be a source for isolation of therapeutic compounds against the dengue-2 virus.

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Prolonged detection of dengue virus RNA in the semen of a man returning from Thailand to Italy, January 2018

Eurosurveillance ◽

10.2807/1560-7917.es.2018.23.18.18-00197 ◽

2018 ◽

Vol 23 (18) ◽

Cited By ~ 16

Author(s):

Eleonora Lalle ◽

Francesca Colavita ◽

Marco Iannetta ◽

Saba Gebremeskel Teklè ◽

Fabrizio Carletti ◽

...

Keyword(s):

Dengue Virus ◽

Viral Replication ◽

Dengue Fever ◽

Symptom Onset ◽

Sexual Transmission ◽

Negative Strand ◽

Cellular Fraction ◽

Virus Rna

This study reports the presence of dengue virus RNA in longitudinally collected semen samples of a previously healthy Caucasian man, returning to Italy from Thailand with primary dengue fever, up to 37 days post-symptom onset, when viraemia and viruria were undetectable. This finding, coupled with the evidence of dengue virus negative-strand RNA, an indirect marker of ongoing viral replication, in the cellular fraction of semen, indicates a need to further investigate possible sexual transmission.

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