Abstract
Chikungunya virus (CHIKV) non-structural protein 2 (nsP2) is a multifunctional enzyme essential for viral replication and host immune evasion. While nsP2's role in mammalian infection is well-established, its interactions within mosquito vectors remain poorly characterized. Here, we performed a comparative interactome analysis using co-immunoprecipitation coupled with liquid chromatography-tandem mass spectrometry to identify nsP2 host protein interactions in human hepatoma-derived Huh7 cells and Aedes albopictus U4.4 cells. We identified 56 high-confidence nsP2 interactors in Huh7 cells and 46 in U4.4 cells, with pathway enrichment analysis revealing convergent targeting of translation machinery, proteasomal degradation, and metabolic pathways in both systems. Protein-protein interaction networks demonstrated distinct functional clustering, with mammalian networks emphasizing metabolic reprogramming and mitochondrial function, while mosquito networks prioritized translation and oxidative stress responses. Comparative analysis identified six conserved interactors across both cell types, highlighting fundamental host dependencies exploited by CHIKV across vertebrate and invertebrate hosts, providing insights for developing broad-spectrum antiviral strategies targeting virus-host interfaces critical for replication in both mammalian hosts and mosquito vectors. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13337-025-00941-x.