Identifying Interactions Between Chikungunya Virus And Aedes Aegypti Salivary Gland Proteins

Alfred Jiang

This thesis is restricted to Yale network users only. It will be made publicly available on 08/28/2021


Chikungunya virus (CHIKV), an emerging threat to global health, has been responsible for extensive arboviral epidemics in Southeast Asia and the Americas in recent years. In order to develop effective therapeutics and preventative measures, we must first gain a deeper understanding of both the host and vector factors that define the pathogen’s life cycle and transmissibility. Currently, the entry receptor and potential co-factors that facilitate CHIKV infection in the arthropod vector are unknown, highlighting a critical knowledge gap in our understanding of the viral life cycle. Furthermore, while secreted mosquito salivary proteins have been shown to modulate host infection through physical interactions with the virion for dengue and West Nile virus, such interactions have not been explored for CHIKV. In this study, we explore three different approaches for screening interactions between CHIKV and proteins expressed by the Ae. aegypti vector. Specifically, we detail the use of a yeast display library constructed from the salivary glands of Ae. aegypti as well as co-immunoprecipitation protocols using Ae. aegypti saliva and the plasma membrane fraction of Aag-2 cells. By using these techniques, we identified three candidates that may interact with the envelope (E2) protein of CHIKV: tryptase-like protein (LOC5575351), uncharacterized protein (LOC5578922), and odorant-binding protein 99b (LOC5564586).