Complete Genome Sequences of Four Serotypes of Dengue Virus Prototype Continuously Maintained in the Laboratory

Chin-inmanu, Kwanrutai; Suttitheptumrong, Aroonroong; Sangsrakru, Duangjai; Mairiang, Dumrong; Tangphatsornruang, Sithichoke; Malasit, Prida; Suriyaphol, Prapat

doi:10.1128/mra.00199-19

“…To examine whether the observed effects were specific to DENV2-NGC, mRNA depletion experiments were repeated in cells infected with Thailand strain DENV2-16681, which has distinct mutations from the DENV2-NGC virus [ 27 ]. Results in S1C Fig show that depletion of Sec61A1 or Loqs also diminished DENV2-16681 RNA abundances.…”

Section: Resultsmentioning

confidence: 99%

Loquacious modulates flaviviral RNA replication in mosquito cells

Shwetha

¹

,

Weng

²

,

Ooi

³

et al. 2022

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Arthropod-borne viruses infect both mosquito and mammalian hosts. While much is known about virus-host interactions that modulate viral gene expression in their mammalian host, much less is known about the interactions that involve inhibition, subversion or avoidance strategies in the mosquito host. A novel RNA-Protein interaction detection assay was used to detect proteins that directly or indirectly bind to dengue viral genomes in infected mosquito cells. Membrane-associated mosquito proteins Sec61A1 and Loquacious (Loqs) were found to be in complex with the viral RNA. Depletion analysis demonstrated that both Sec61A1 and Loqs have pro-viral functions in the dengue viral infectious cycle. Co-localization and pull-down assays showed that Loqs interacts with viral protein NS3 and both full-length and subgenomic viral RNAs. While Loqs coats the entire positive-stranded viral RNA, it binds selectively to the 3’ end of the negative-strand of the viral genome. In-depth analyses showed that the absence of Loqs did not affect translation or turnover of the viral RNA but modulated viral replication. Loqs also displayed pro-viral functions for several flaviviruses in infected mosquito cells, suggesting a conserved role for Loqs in flavivirus-infected mosquito cells.

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“…To examine whether the observed effects were specific to DENV2-NGC, mRNA depletion experiments were repeated in cells infected with Thailand strain DENV2-16681, which has distinct mutations from the DENV2-NGC virus [24]. Results in Fig.…”

Section: Resultsmentioning

confidence: 99%

Loquacious Modulates Flaviviral RNA Replication in Mosquito Cells

Shwetha

¹

,

Weng

²

,

Ooi

³

et al. 2021

Preprint

0

View full text Add to dashboard Cite

Arthropod-borne viruses infect both mosquito and mammalian hosts. While much is known about virus-host interactions that modulate viral gene expression in their mammalian host, much less is known about the interactions that involve inhibition, subversion or avoidance strategies in the mosquito host. A novel RNA-Protein interaction detection assay was used to detect proteins that directly or indirectly bind to dengue viral genomes in infected mosquito cells. Membrane-associated mosquito proteins SEC61A1 and Loquacious (Loqs) were found to be in complex with the viral RNA. Depletion analysis demonstrated that both SEC61A1 and Loqs have pro-viral functions in the dengue viral infectious cycle. Co-localization and pull-down assays showed that Loqs interacts with viral protein NS3 and both full-length and subgenomic viral RNAs. While Loqs coats the entire positive-stranded viral RNA, it binds selectively to the 3’ end of the negative-strand of the viral genome. In-depth analyses showed that the absence of Loqs did not affect translation or turnover of the viral RNA but modulated viral replication. Loqs also displayed pro-viral functions for several flaviviruses in infected mosquito cells, suggesting a conserved role for Loqs in flavivirus-infected mosquito cells.Author SummaryThere is a wealth of information that dictates virus-host interactions in flavivirus-infected mammalian cells, yet there is only sparse information on the mechanisms that modulate viral gene expression in the mosquito host. Using a novel RNA-protein detection assay, the interactions of SEC61A1 and Loqs with the dengue viral genome were found to have proviral functions in infected mosquito cells. In particular, Loqs forms complexes with the positive-strand of the viral RNA and the very 3’ end of the negative-strand viral RNA. Further analyses showed that Loqs modulates viral RNA replication of dengue virus and gene amplification of several other flaviviral genomes. These findings argue that Loqs is an essential proviral host factor in mosquitos.

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“…DENV reference strains are widely used for virologic studies and have been cultured under different laboratory conditions, which results in the accumulation of genetic variations. 33 These variations do not faithfully reflect the biological behavior of natural DENV populations in Colombia. On the other hand, the immune response and the history of viral infections of the patient have been described as factors that influence the population dynamics of DENV.…”

Section: Discussionmentioning

confidence: 99%