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Acta Biológica Colombiana

versão impressa ISSN 0120-548X

Resumo

BUENO ANGARITA, Claudia Liliana et al. Assembly and release of Dengue virus: Role of Alix protein. Acta biol.Colomb. [online]. 2019, vol.24, n.3, pp.509-519. ISSN 0120-548X.  https://doi.org/10.15446/abc.v24n3.79321.

Since the finding that HIV recruits cellular ESCRT (endosomal sorting complexes required for transport) machinery to accomplish viral budding, this strategy has emerged as an escape route for enveloped viruses also. The work aimed to explore the participation of the cellular protein Alix (a human protein that acts as an adapter in the ESCRT pathway) on the transcription, protein expression, assembly and release of Dengue virus (DENV), and explore for its potential interaction with the viral protein NS3. To this purpose, A549 cells were infected with DENV2 and treated with small interfering RNAs (siRNA) to generate an Alix stable knockdown cells line. Also, an A549 cells line expressing a histidine-tagged NS3 protein was obtained. Both cells lines were used in immunoprecipitation and pull-down assays to assess the interaction between NS3 and Alix. The results showed that Alix knockdown had no effect on viral transcription or viral protein expression but influenced the assembly and release of DENV2 negatively. Finally, pull-down assays revealed the interaction between NS3 and Alix. The finding of an Alix participation in the production of DENV2 and its interaction with NS3 provides a potential target for the design of control/inhibition strategies against DENV spread.

Palavras-chave : Flavivirus; ESCRT machinery; serine endopeptidase; virus physiological processes.

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