UWEC CERCA 2026

RIG-I-like receptors (RLRs) are important cytosolic sensors that detect and respond to viral dsRNA during an infection. This family is characterized by the conserved RIG-I-like helicase domain that binds dsRNA and hydrolyzes ATP. Many viruses have evolved mechanisms to evade or suppress this mechanism, including the expression of Viral Suppressors of RNA Sensing (VSR) proteins. Although the role of RLR signaling is well-studied, specific VSR-RLR protein interactions are not fully characterized. In this project, we utilized the Yeast Two-Hybrid method to identify whether candidate VSRs interact with the RLRs RIG-I, MDA5, LGP2, and Dicer. Yeast are transformed with a pair of plasmids containing the split halves of the yeast GAL4 transcription factor. The activation domain (AD) is fused with one of the RLRs (‘bait’) while the DNA-binding domain (DNA-BD) is fused with a VSR (‘prey’). After co-transformation and culturing on selective media, yeast can only grow if the bait and prey interact. This method allows us to screen many VSR-RLR combinations to determine if VSRs are specific inhibitors of one RLR or general inhibitors of the family. Future work will determine if any detected interactions are dependent on the RIG-I-like helicase domain. Overall, this project provides insight into virus-host interactions during infection and the important of RLRs to innate immunity.