The dengue virus non-structural protein 1 (NS1) is secreted efficiently from infected mosquito cells

Alcalá, Ana C.; Medina, Fernando; González‐Robles, Arturo; Salazar‐Villatoro, Lizbeth; Fragoso‐Soriano, Rogelio; Vásquez, Carlos; Cervantes-Salazar, Margot; Ángel, Rosa M. del; Ludert, Juan E.

doi:10.1016/j.virol.2015.11.020

Cited by 40 publications

(36 citation statements)

References 35 publications

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“…These results are consistent with a study in which mutation of Asn130 resulted in a loss of hexameric NS1 and an increase in the formation of higher-order NS1 oligomers (34). Indeed, our data agree with recent studies showing that NS1 secreted from DENV-infected C6/36 cells is hexameric in nature (51). …”

Section: Discussionsupporting

confidence: 93%

Secreted NS1 Protects Dengue Virus from Mannose-Binding Lectin–Mediated Neutralization

Thiemmeca

Tamdet

Punyadee

et al. 2016

The Journal of Immunology

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Flavivirus nonstructural protein 1 (NS1) is a unique secreted non-structural glycoprotein. Although it is absent from the flavivirus virion, intracellular and extracellular forms of NS1 have essential roles in viral replication and the pathogenesis of infection. The fate of NS1 in insect cells has been more controversial, with some reports suggesting it is exclusively cell-associated. Here, we confirm NS1 secretion from cells of insect origin and characterize its physical, biochemical, and functional properties in the context of dengue virus (DENV) infection. Unlike mammalian cell-derived NS1, which displays both high mannose and complex type N-linked glycans, soluble NS1 secreted from DENV-infected insect cells contains only high mannose glycans. Insect cell-derived secreted NS1 also has different physical properties including smaller and more heterogeneous sizes, and the formation of less stable NS1 hexamers. Both mammalian and insect cell-derived NS1 bind to complement proteins C1s, C4 and C4 binding protein as well as to a novel partner, mannose-binding lectin (MBL). Binding of NS1 to MBL protects DENV virus against MBL-mediated neutralization by the lectin pathway of complement activation. As we detected secreted NS1 and DENV virus together in the saliva of infected Aedes aegypti mosquitoes, these findings suggest a mechanism of viral immune evasion at the very earliest phase of infection.

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Section: Discussionsupporting

confidence: 93%

Secreted NS1 Protects Dengue Virus from Mannose-Binding Lectin–Mediated Neutralization

Thiemmeca

Tamdet

Punyadee

et al. 2016

The Journal of Immunology

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“…In addition, we found smaller particles with a diameter of 13.1±2.1 nm, which had either electron-translucent or electron-dense cores. Presumably, they could represent either: a) glycoprotein aggregates, similar to the "classic" flavivirus nonstructural protein 1, which is secreted from infected cells (36,37), b) protein structures appearing after long-term persistence, c) virions with incomplete genome, or d) other unknown structures present in IRE/CTVM19 cells. Further investigations are needed to clarify this point.…”

Section: Discussionmentioning

confidence: 99%

First isolation and characterisation of Alongshan virus in Russia

Kholodilov

Litov

Klimentov

et al. 2019

Preprint

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AbstractIn recent decades, many new flavi-like viruses have been discovered predominantly in different invertebrates and, as was recently shown, some of them may cause disease in humans. The Jingmen tick virus (JMTV) group holds a special place among flavi-like viruses because, in contrast to the “classic” flaviviruses and other flavi-like viruses, they have a segmented ssRNA(+) genome. Two segments of the JMTV genome have homology with regions of the flavivirus genome encoding polymerase and helicase-protease proteins. JMTVs have several open reading frames (ORF) in segments encoding glycoprotein(s) and capsid protein and these ORF are specific only to them. JMTVs greatly differ in virion size.We isolated three strains of Alongshan virus (ALSV), which is a representative of the JMTV group, from adult Ixodes persulcatus ticks collected in two geographically-separated Russian regions in the tick cell line IRE/CTVM19. One of the strains persisted in the IRE/CTVM19 cells without cytopathic effect for three years. Most virions purified from tick cells were spherical with a diameter of approximately 40.5 nm. In addition, we found smaller particles of approximately 13.1 nm in diameter. We obtained full genome sequences of all four segments of two of the isolated ALSV strains, and partial sequences of one segment from the third strain. Phylogenetic analysis on genome segment 2 of the JMTV group clustered our novel strains with other ALSV strains. We found evidence for the existence of a novel upstream ORF in the glycoprotein-coding segment of ALSV and other members of the JMTV group.Significance StatementWe isolated three strains of Alongshan virus (ALSV) from adult Ixodes persulcatus ticks from two geographically separate areas of Russia in the Ixodes ricinus tick cell line IRE/CTVM19. One of the strains persisted in the IRE/CTVM19 cells without cytopathic effect for three years. Our study confirmed the value of tick cell lines in virus isolation and maintenance of persistent infection. The majority of virions of the ALSV strain Miass527 were enveloped spherical particles with a diameter of 40.5±3.7 nm. We found evidence for the existence of a novel upstream ORF in the glycoprotein-coding segment of ALSV and other members of the Jingmen tick virus group.

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“…Recent studies have shown that the DENV NS1 protein was not only secreted from vertebrate cells, but that was also efficiently secreted from mosquito cells lines [7,8]. The secretion of NS1 in vertebrate cells follows the classical Golgi-pathway [9].…”

Section: Introductionmentioning

confidence: 99%

The dengue virus non-structural protein 1 (NS1) is secreted from mosquito cells in association with the intracellular cholesterol transporter chaperone caveolin complex

Ramirez

Ludert

2018

Preprint

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11 12 Keywords: 13 dengue virus; zika virus; yellow fever virus; flavivirus; NS1; caveolin-1; chaperone caveolin 14 complex; viral protein trafficking; unconventional secretion; mosquito cells 15 16 17 ABSTRACT 18 Dengue virus (DENV) is a mosquito-borne virus of the family Flaviviridae. The RNA viral genome 19 encodes for a polyprotein that is co-translationally processed into three structural proteins and . CC-BY 4.0 International license is made available under aThe copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 2 20 seven non-structural proteins. The non-structural protein 1 (NS1) is a multifunctional viral protein 21 actively secreted in vertebrate and mosquito cells during DENV infection. In mosquito cells, NS1 22 is secreted in a caveolin-1 (CAV-1) dependent manner by an unconventional pathway. The 23 caveolin chaperone complex (CCC) is a cytoplasmic complex formed by caveolin-1 and the 24 chaperones FKBP52, Cy40 and CyA which is responsible for cholesterol traffic inside the cell. 25 In this work, we demonstrate that in infected mosquito cells, DENV NS1 is secreted by an early 26 and unconventional route that bypasses the Golgi apparatus in close association with the CCC.27 Treatment of mosquito cells with classic secretion inhibitors such as brefeldin A, golgicide A and 28 Fli-06 showed no effect on NS1 secretion, but significant reductions in recombinant luciferase 29 secretion and virion release. Silencing the expression of CAV1, FKBP52 with siRNAs or the 30 inhibition of CyA by cyclosporine A resulted in significant decrease in NS1 secretion without 31 affecting virion release. Using co-localization, co-inmunoprecipitation and proximity ligation 32 assays, NS1 was found to co-localize and interact with all the protein components of the CCC 33 in mosquito infected cells. In addition, CAV-1 and FKBP52 expression was found augmented in 34 DENV infected cells. Finally, the treatment of ZIKV infected mosquito cells with brefeldin A and 35 golgicide A showed no effect on NS1 secretion, while affecting virion release. ZIKV NS1 was 36 also found to co-localize with CAV-1 in infected mosquito cells. These results suggest that in 37 mosquito cells, ZIKV NS1 follows the same secretory pathway observed for DENV NS1. The 38 association of NS1 with the cholesterol transporter CCC agrees with the lipoprotein nature of 39 secreted hexameric NS1. 40 41 AUTHOR SUMMARY . CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 3 42 Dengue protein NS1 is secreted in infected mosquito and vertebrate cells. In humans, secreted 43 NS1 have been associated with pathogenesis. In mosquito cells, NS1 follows an unconventional 44 secretion pathway that is dependent on Caveolin-1. This work shows that in mosquito cells, NS1 45 secretion is associated to the chaperone caveolin complex, a...

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The dengue virus non-structural protein 1 (NS1) is secreted efficiently from infected mosquito cells

Cited by 40 publications

References 35 publications

Secreted NS1 Protects Dengue Virus from Mannose-Binding Lectin–Mediated Neutralization

Secreted NS1 Protects Dengue Virus from Mannose-Binding Lectin–Mediated Neutralization

First isolation and characterisation of Alongshan virus in Russia

The dengue virus non-structural protein 1 (NS1) is secreted from mosquito cells in association with the intracellular cholesterol transporter chaperone caveolin complex

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