Instability of the NS1 Glycoprotein from La Reunion 2018 Dengue 2 Virus (Cosmopolitan-1 Genotype) in Huh7 Cells Is Due to Lysine Residues on Positions 272 and 324

Ogire, Eva; Diaz, Olivier; Vidalain, Pierre‐Olivier; Lotteau, Vincent; Desprès, Philippe; Roche, Marjolaine

doi:10.3390/ijms22041951

Cited by 4 publications

(3 citation statements)

References 47 publications

(67 reference statements)

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“…Although further studies are necessary, the in vivo differences might have been due, at least partly, to the significant impact of NS1 proteins in increasing the pathogenicity. Also, the lysine residues at positions 272 and 324 have been reported to contribute to the instability of the NS1 glycoprotein in human hepatocyte HuH7 cells [35]. A lysine residue was almost always found at position 272 in the Asian-I viruses (S5 Fig), while arginine was found at position 272 in the Cosmopolitan virus used in this experiment.…”

Section: Plos Neglected Tropical Diseasesmentioning

confidence: 55%

Genetic regions affecting the replication and pathogenicity of dengue virus type 2

Samune,

Saito,

Sasaki

et al. 2024

PLoS Negl Trop Dis

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Dengue is a mosquito-borne disease that has spread to over 100 countries. Its symptoms vary from the relatively mild acute febrile illness called dengue fever to the much more severe dengue shock syndrome. Dengue is caused by dengue virus (DENV), which belongs to the Flavivirus genus of the family Flaviviridae. There are four serotypes of DENV, i.e., DENV1 to DENV4, and each serotype is divided into distinct genotypes. Thailand is an endemic area where all four serotypes of DENV co-circulate. Genome sequencing of the DENV2 that was isolated in Thailand in 2016 and 2017 revealed the emergence of the Cosmopolitan genotype and its co-circulation with the Asian-I genotype. However, it was unclear whether different genotypes have different levels of viral replication and pathogenicity. Focus-forming assay (FFA) results showed that clinical isolates of these genotypes differed in focus size and proliferative capacity. Using circular polymerase extension reaction, we generated parental and chimeric viruses with swapped genes between these two DENV2 genotypes, and compared their focus sizes and infectivity titers using FFA. The results showed that the focus size was larger when the structural proteins and/or non-structural NS1-NS2B proteins were derived from the Cosmopolitan virus. The infectious titers were consistent with the focus sizes. Single-round infectious particle assay results confirmed that chimeric viruses with Cosmopolitan type structural proteins, particularly prM/E, had significantly increased luciferase activity. Replicon assay results showed that Cosmopolitan NS1-NS2B proteins had increased reporter gene expression levels. Furthermore, in interferon-receptor knock-out mice, viruses with Cosmopolitan structural and NS1-NS2B proteins had higher titers in the blood, and caused critical disease courses. These results suggested that differences in the sequences within the structural and NS1-NS2B proteins may be responsible for the differences in replication, pathogenicity, and infectivity between the Asian-I and Cosmopolitan viruses.

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Section: Plos Neglected Tropical Diseasesmentioning

confidence: 55%

Genetic regions affecting the replication and pathogenicity of dengue virus type 2

Samune,

Saito,

Sasaki

et al. 2024

PLoS Negl Trop Dis

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“…Expression of recombinant RUN-18 E protein was assessed in A549 cells transfected with pcDNA3/RUN-18 prME for 18 h. A plasmid pcDNA3/RUN-18 NS1 encoding recombinant soluble NS1 protein from RUN-18 was used as a plasmid control since protein expression results in a weak effect on cell viability [29]. Expression of recombinant RUN-18 NS1 in A549 cells transfected with pcDNA3/RUN-18 NS1 was verified by FACS analysis and immunoblot assay (Fig.S1, available in the online version of this article).…”

Section: Resultsmentioning

confidence: 99%

Dengue virus M and E proteins belonging to genotype II (Cosmopolitan) of serotype 2 are influenced by the nature of M residue 36

Decotter

Desprès

Gadéa

2023

Journal of General Virology

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Mosquito-borne dengue disease is caused by the dengue virus serotype-1 to serotype-4. The contemporary dengue outbreaks in the southwestern Indian ocean coincided with the widespread of dengue virus serotype 2 genotype II (Cosmopolitan), including epidemic viral strains DES-14 and RUN-18 isolated in Dar es Salaam (Tanzania) in 2014 and La Reunion Island (France) in 2018, respectively. Heterodimeric interaction between prM (intracellular precursor of surface structural M protein) and envelope E proteins is required during the initial stage of dengue virus assembly. Amino acid 127 of DES-14 prM protein (equivalent to M36) has been identified as an infrequent valine whereas RUN-18 has a common isoleucine. In the present study, we examined the effect of M-I36V mutation on the expression of a recombinant RUN-18 E protein co-expressed with prM in human epithelial A549 cells. The M ectodomain of dengue virus serotype 2 embeds a pro-apoptotic peptide referred as D2AMP. The impact of M-I36V mutation on the death-promoting capability of D2AMP was assessed in A549 cells. We showed that valine at position M36 affects expression of recombinant RUN-18 E protein and potentiates apoptosis-inducing activity of D2AMP. We propose that the nature of M residue 36 influences the virological characteristics of dengue 2 M and E proteins belonging to genotype II that contributes to global dengue burden.

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“…Expression of recombinant RUN-18 E protein was assessed in A549 cells transfected with pcDNA3/RUN-18 prME for 18 h. A plasmid pcDNA3/RUN-18 NS1 encoding recombinant soluble NS1 protein from RUN-18 was used as a plasmid control since protein expression results in a weak effect on cell viability [21]. Expression of recombinant RUN-18 NS1 in A549 cells transfected with pcDNA3/RUN-18 NS1 was verified by FACS analysis and immunoblot assay (Figure S1).…”

Section: Characterization Of Recombinant Run-18 E Protein Co-expresse...mentioning

confidence: 99%

The hydrophobic nature of dengue 2 M residue 36 influences E protein and ApoptoM peptide

Decotter

Després

Gadéa³

2023

Preprint

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The recent epidemics of dengue in South West Indian Ocean coincided with the emergence of Cosmopolitan dengue virus type 2, including viral strains DES-14 in Tanzania and then RUN-18 in La Reunion. The initial step of dengue virus assembly is the formation of heterodimers between prM and E proteins where prM acts as a chaperone for E. During dengue virus maturation, prM is cleaved into membrane protein M which embeds a pro-apoptotic peptide consisting of residues 31/41 and referred as ApoptoM. An infrequent valine at position M-36 was found in DES-14 whereas RUN-18 bears a common isoleucine. Here, we investigated whether Ile-to-Val substitution at position M-36 may have an impact on RUN-18 E expression and cell-death promoting capability of RUN-18 ApoptoM. Using recombinant RUN-18 envelope proteins expressed in human epithelial A549 cells, we showed that Ile-to-Val but not Ile-to-Ala substitution affects the behavior E protein reducing the cytotoxicity of prM and E proteins. The substitution of isoleucine by valine at position M-36 leads to increase the apoptosis-inducing activity of ApoptoM. Our data highlight that hydrophobic nature of amino-acid residue in position 36 of dengue virus M protein influences E protein expression and death-promoting activity of ApoptoM opens up important perspectives in the development of effective live-attenuated DENV vaccines.

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Instability of the NS1 Glycoprotein from La Reunion 2018 Dengue 2 Virus (Cosmopolitan-1 Genotype) in Huh7 Cells Is Due to Lysine Residues on Positions 272 and 324

Cited by 4 publications

References 47 publications

Genetic regions affecting the replication and pathogenicity of dengue virus type 2

Genetic regions affecting the replication and pathogenicity of dengue virus type 2

Dengue virus M and E proteins belonging to genotype II (Cosmopolitan) of serotype 2 are influenced by the nature of M residue 36

The hydrophobic nature of dengue 2 M residue 36 influences E protein and ApoptoM peptide

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