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

Abstract

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, Aedes aegypti. 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.

Protein-protein interactions between A. aegypti midgut and dengue virus 2: two-hybrid screens using the midgut cDNA library

“…In another study, Mairiang et al identified several mosquito protein interacting partners of DENV-2 C, prM, NS3, NS4A, NS4B, and NS5 proteins [23]. Interestingly, the study identified similar proteins that were also identified in our study.…”

“…Interestingly, the study identified similar proteins that were also identified in our study. These include human ribosomal protein and mosquito carboxypeptidase, which were suggested to interact with the DENV C protein [23]. However, although most of the earlier studies have identified putative interacting partners of DENV proteins, very limited information is available on mosquito protein interactors of DENV E and NS1 proteins.…”

“…Carboxypeptidase, a well-known hydrolytic enzyme involved in C-terminal peptide cleavage, was also found to be highly regulated after blood meal [53]. In mosquito, carboxypeptidase has not only been found to be involved in sexual development of malarial protozoan parasites [54], but also in the interaction between carboxypeptidase and DENV capsid protein in the salivary gland [23] and the midgut cells [55]. In addition, carboxypeptidase D has been continuously proven to be a receptor for duck hepatitis B virus [56].…”

“…To date, very few studies have been conducted to determine the interactome between DENV proteins and mosquito cellular proteins using sensitive and reliable protein interaction assays [22,23], with some predictions reported through a computational approach [24]. Hence, the aim of this study was to identify adult A. aegypti mosquito midgut proteins interacting with DENV-2 viral proteins, using an improved yeast twohybrid (Y2H) screening system, a high-throughput screening assay that allows identification of genuine protein interacting partners in vivo.…”

Development of viable TAP-tagged dengue virus for investigation of host–virus interactions in viral replication

“…A number of approaches have been used to identify NS5-interacting partners (Carpp et al, 2014;Doolittle & Gomez, 2011;Khadka et al, 2011;Le Breton et al, 2011;Mairiang et al, 2013). The yeast two-hybrid system (Y2H) offers the advantage of rapid large-scale screening of protein pairs.…”