Abstract
Dengue virus (DENV) is a flavivirus that causes marked human morbidity and mortality worldwide, and is transmitted to humans by Aedes aegypti mosquitoes. Habitat expansion of Aedes, mainly due to climate change and increasing overlap between urban and wild habitats, places nearly half of the world’s population at risk for DENV infection. After a bloodmeal from a DENV-infected host, the virus enters the mosquito midgut. Next, the virus migrates to, and replicates in, other tissues, like salivary glands. Successful viral transmission occurs when the infected mosquito takes another blood meal on a susceptible host and DENV is released from the salivary gland via saliva into the skin. During viral dissemination in the mosquito and transmission to a new mammalian host, DENV interacts with a variety of vector proteins, which are uniquely important during each phase of the viral cycle. Our study focuses on the interaction between DENV particles and protein components in the A. aegypti vector. We performed a mass spectrometry assay where we identified a set of A. aegypti salivary gland proteins which potentially interact with the DENV virion. Using dsRNA to silence gene expression, we analyzed the role of these proteins in viral infectivity. Two of these candidates, a synaptosomal-associated protein (AeSNAP) and a calcium transporter ATPase (ATPase) appear to play a role in viral replication both in vitro and in vivo, observing a ubiquitous expression of these proteins in the mosquito. These findings suggest that AeSNAP plays a protective role during DENV infection of mosquitoes and that ATPase protein is required for DENV during amplification within the vector.
Highlights
Dengue is a major public health threat in tropical and subtropical areas, and as climate change and urbanization continues, the illness may spread to other locations across the globe [1]
To identify A. aegypti salivary gland components that potentially interact with Dengue virus (DENV) virions, we utilized gradient sucrose purification of DENV virions that were pre-incubated with mosquito salivary gland extracts (SGE)
DENV virions alone were used as control, allowing us to identify vector peptides that were only detected in samples containing SGE
Summary
Dengue is a major public health threat in tropical and subtropical areas, and as climate change and urbanization continues, the illness may spread to other locations across the globe [1]. According to reports from the World Health Organization, before 1970 only nine countries experienced outbreaks of severe dengue. More attention is required to develop countermeasures to address this expansion, fed by processes including global warming, unprecedented human mobility, rapid urban population growth, and large-scale changes in ecosystems [2,3,4,5]. After taking a viremic blood meal, DENV establishes infection in the midgut. Upon establishing a successful infection, the virus disseminates systemically through the hemolymph where it can invade secondary tissues, such as the salivary glands [6]. Replication in the salivary glands leads to virion release into the saliva, the last step prior to virus transmission to the human host [7]
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