7 research outputs found

    Identification of New Protein Interactions between Dengue Fever Virus and Its Hosts, Human and Mosquito

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    <div><p>The four divergent serotypes of dengue virus are the causative agents of dengue fever, dengue hemorrhagic fever and dengue shock syndrome. About two-fifths of the world's population live in areas where dengue is prevalent, and thousands of deaths are caused by the viruses every year. Dengue virus is transmitted from one person to another primarily by the yellow fever mosquito, <em>Aedes aegypti</em>. Recent studies have begun to define how the dengue viral proteins interact with host proteins to mediate viral replication and pathogenesis. A combined analysis of these studies, however, suggests that many virus-host protein interactions remain to be identified, especially for the mosquito host. In this study, we used high-throughput yeast two-hybrid screening to identify mosquito and human proteins that physically interact with dengue proteins. We tested each identified host protein against the proteins from all four serotypes of dengue to identify interactions that are conserved across serotypes. We further confirmed many of the interactions using co-affinity purification assays. As in other large-scale screens, we identified some previously detected interactions and many new ones, moving us closer to a complete host – dengue protein interactome. To help summarize and prioritize the data for further study, we combined our interactions with other published data and identified a subset of the host-dengue interactions that are now supported by multiple forms of evidence. These data should be useful for understanding the interplay between dengue and its hosts and may provide candidates for drug targets and vector control strategies.</p> </div

    Dengue-host interactions supported by multiple forms of evidence.

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    <p>(A) Dengue-human interactome. (B) Dengue-mosquito interactome. Pink nodes represent host proteins. Green nodes represent dengue proteins. Red edges represent PPI with conserved interologs. Additional details are available in Cytoscape files in supplemental data (Data S1).</p

    Dengue – host protein networks derived from two-hybrid screens and co-AP assays in this study.

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    <p>(A) Human-dengue interaction map. (B) Mosquito-dengue interaction map. Edges represent protein-protein interactions. Green nodes are dengue proteins, yellow nodes are host proteins, and blue nodes are host proteins found in both the human and mosquito maps. Red edges represent protein-protein interactions universally detected for all four serotypes. Blue edges represent protein-protein interactions confirmed by co-AP assays. Green edges represent the universal interactions that were confirmed by co-AP assays. Additional details are available in Cytoscape files in supplemental data (Data S1).</p

    Dengue virus proteins.

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    <p>(A) The dengue polyprotein in the ER membrane prior to processing. (B) The coding regions for the fourteen dengue virus proteins and partial peptides shown were separately cloned into yeast two-hybrid plasmids.</p

    Examples of co-AP results.

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    <p>Host proteins were fused to a myc-tag while dengue proteins were fused to an NTAP-tag. The fusion proteins were expressed in S2R+ cells. NTAP-dengue proteins were purified from cell lysates, and then host proteins were detected with Ξ±-myc. (A) An Ξ±-myc immunoblot of cell lysates shows expression of mosquito and human proteins. (B) An Ξ±-myc immunoblot of NTAP-tag affinity-purified samples. Additional co-AP results are in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053535#pone.0053535.s014" target="_blank">Figure S3</a>.</p
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