Putative dengue virus receptors from mosquito cells

Abstract

Dengue viruses are arthropod-borne, single-stranded RNA viruses. Aëdes aegypti and Aëdes albopictus are the principal vectors. In order to understand the molecular basis of dengue virus infections we explored the biochemical identity of dengue-2 (DEN-2) virus receptors in the Aëdes albopictus-derived cell line C6/36. We show here that DEN-2 interacts with two major polypeptides of 80 and 67 kDa. Polyclonal anti-C6/36 membrane antibodies block DEN-2 binding to intact C6/36 monolayers as well as to membrane extracts. Our results strongly suggest that the identified polypeptides are part of the DEN-2 receptors.

Proteomic Identification of Dengue Virus Binding Proteins inAedes aegyptiMosquitoes andAedes albopictusCells

“…NHS-coupled biotinyl compounds have been used to label cell surface proteins previously [8]. The procedure to biotinylate DENV particles was as follow.…”

“…Dengue virus attaches to the host epithelial cell receptors protein E-mediated [8, 9] and enters the cell mainly via this receptor by clathrin-dependent endocytosis [10–13]. …”

“…The apparent molecular weights described for these proteins are between 20 to 40 kDa and 57 to 130 kDa in size and bind dengue virus particles in vitro [8, 20–24]. In addition, Mercado-Curiel et al [23] reported that specific antibodies against the membrane proteins R67 and R80 inhibited infection of C6/36 cells.…”

A DNA Microarray-Based Assay to Detect Dual Infection with Two Dengue Virus Serotypes

“…DENV (genus Flavivirus , family Flaviviridae ) is mosquito-borne and mainly transmitted by Aedes ( Stegomyia ) aegypti (L.) and Aedes ( Stegomyia ) albopictus ( Skuse ) ( Diptera : Culicidae ), which are infected through receptors in the midgut [6–8]. This virus has a positive-strand RNA genome of approximately 11,000 bases, which encodes three structural and seven non-structural proteins.…”

A gradient-free method for the purification of infective dengue virus for protein-level investigations

“…Dengue virus (DENV) is the causative agent of Dengue Fever (DF), Dengue Hemorrhagic Fever (DHF), and Dengue Shock Syndrome (DSS) which conservatively affect 100 million people annually (Laughlin et al, 2012). Dengue virus and other flaviviruses are commonly studied after propagation in Aedes albopictus C6/36 cells, (Igarashi, 1978; Sakoonwatanyoo et al, 2006) and are purified for protein-level analysis; including the determination of protein–protein interactions (Muñoz et al, 1998), structural studies (Kuhn et al, 2002), and diagnosis (Peeling et al, 2010). Disease-relevant viruses known to infect the Aedes mosquitoes include DENV, West Nile virus (Colpitts et al, 2012), Zika virus (Grard et al, 2014), Yellow Fever virus (Hanley et al, 2013), and Chikungunya virus (Li et al, 2012).…”