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White spot syndrome virus (WSSV) is one major pathogen in shrimp aquaculture. WSSV ORF403 is predicted to encode a protein of 641 amino acids, which contains a C3H2C2 RING structure. In the presence of an E2 conjugating enzyme from shrimp, WSSV403 can ubiquitinate itself  in vitro , indicating it can function as a viral E3 ligase. Besides, WSSV403 E3 ligase can be activated by a series of E2 variants. Based on RT-PCR and Real time PCR, we detected transcription of WSSV403 in the commercial specific-pathogen-free (SPF) shrimp, suggesting its role as a latency-associated gene. Identified in yeast two-hybrid screening and verified by pull-down assays, WSSV403 is able to bind to a shrimp protein phosphatase (PPs), which was characterized before as an interaction partner for another latent protein WSSV427. Our studies suggest that WSSV403 is a regulator of latency state of WSSV by virtue of its E3 ligase function.

Identification and characterization of a new E3 ubiquitin ligase in white spot syndrome virus involved in virus latency