Regulation of Epstein-Barr Virus Minor Capsid Protein BORF1 by TRIM5 alpha

TRIM5 alpha is a host anti-retroviral restriction factor that destroys human immunodeficiency virus (HIV) virions and triggers innate immune signaling. TRIM5 alpha also mediates the autophagic degradation of target proteins via TRIMosome formation. We previously showed that TRIM5 alpha promotes Epstein-Barr virus (EBV) Rta ubiquitination and attenuates EBV lytic progression. In this study, we sought to elucidate whether TRIM5 alpha can interact with and induce the degradation of EBV capsid proteins. Glutathione S-transferase (GST) pulldown and immunoprecipitation assays were conducted to identify interacting proteins, and mutants were generated to investigate key binding domains and ubiquitination sites. Results showed that TRIM5 alpha binds directly with BORF1, an EBV capsid protein with a nuclear localization signal (NLS) that enables the transport of EBV capsid proteins into the host nucleus to facilitate capsid assembly. TRIM5 alpha promotes BORF1 ubiquitination, which requires the surface patch region in the TRIM5 alpha PRY/SPRY domain. TRIM5 alpha expression also decreases the stability of BORF1(6KR), a mutant with all lysine residues mutated to arginine. However, chloroquine treatment restores the stability of BORF1(6KR), suggesting that TRIM5 alpha destabilizes BORF1 via direct recognition of its substrate for autophagic degradation. These results reveal novel insights into the antiviral impact of TRIM5 alpha beyond retroviruses.