Investigation of dengue virus RNA terminal region interactions with human host proteins

Dengue virus (DENV) is a globally important arthropod-borne pathogen responsible for dengue fever, which can range from mild fever to severe hemorrhagic fever and shock syndrome. With no specific antiviral treatments available, understanding the molecular mechanisms of DENV replication and its interactions with host proteins is essential. This study focuses on two interconnected projects that use advanced biophysical techniques to address these challenges. The first project assesses biophysical methods for studying protein-protein interactions (PPIs) in solution, using the sfGFP-anti-GFP nanobody system as a model. Techniques such as microscale thermophoresis, fluorescence correlation spectroscopy, analytical ultracentrifugation, isothermal titration calorimetry, and size exclusion chromatography coupled with multi-angle light scattering were systematically compared. This analysis creates a reliable framework for studying PPIs, which can be applied to protein-RNA and RNA-RNA interactions. The second project examines the interaction between DENV serotype 2 (DENV-2) and the human DEAD-Box Helicase 6 protein (DDX6), a host factor involved in viral replication and assembly. By exploring the molecular interactions between the terminal regions of DENV-2 RNA and DDX6, this work aims to provide mechanistic insights into how DENV hijacks host proteins. To support these studies, DDX6 was expressed recombinantly with a superfolder green fluorescent protein (sfGFP) tag, which facilitates detection in fluorescence-based assays and improves protein yield. Collectively, these findings establish that DDX6 recognizes each structural domain of the DENV-2 3′ UTR independently. Through a robust biophysical framework, we demonstrated this interaction using multiple complementary techniques, extending detailed characterization to a domain that has not been previously studied in depth.

Authors

• Demeler, Aysha K.
• University of Lethbridge. Faculty of Arts and Science

• dengue virus
• RNA terminal region
• human host proteins
• protein-protein interactions
• viral replication

Identifier

hdl:10133/7225

OAI identifier oai:identifier

oai:opus.uleth.ca:10133/7225