Host protein interactome profiling of Akabane virus glycoprotein Gc reveals specific mitochondrial genes regulate viral replication.

Akabane virus (AKAV) is an arthropod-borne orthobunyavirus that poses a significant threat to ruminant health worldwide by causing high fever, stillbirth, and congenital malformations. Although its viral glycoprotein Gc is critical for multiple stages of the viral life cycle, the spectrum of its interactions with host proteins remains uncharacterized. In this study, host proteins interacting with Gc were systematically profiled using co-immunoprecipitation (Co-IP) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total of 441 host proteins were identified as candidate Gc-associating proteins. Comprehensive GO and KEGG enrichment analyses highlighted that these proteins are involved in pathways related to cell cycle regulation, cytoskeletal dynamics, mitochondrial quality control, and ubiquitin-mediated proteolysis. Among the identified candidates, FKBP8, BNIP3, and USP30 were selected for further validation. Co-IP and confocal assays confirmed the interactions of these proteins with Gc. Functional studies demonstrated that overexpression of FKBP8 and BNIP3 significantly enhances AKAV replication, while USP30 acts as a negative regulator of viral proliferation. These findings offer a valuable resource for elucidating AKAV Gc-host interactions and highlight these three mitochondrial proteins as potential targets for AKAV intervention.