WSSV induces Rubicon expression to regulate innate immune response in Penaeus vannamei

Abstract

Rubicon, the RUN domain Beclin-1-interacting and cysteine-rich domain-containing protein plays an important role in facilitating viral replication. In this study, an involvement of P. vannamei Rubicon or PvRUBCN during white spot syndrome virus (WSSV) infection and its roles in regulation of apoptosis and innate immune response were investigated. The full-length coding sequence of PvRUBCN was 3708 bp encoding the protein of 1235 amino acids. PvRUBCN contained three conserved domains including the RUN, the PI3K-binding, and the Rubicon homology domains. PvRUBCN was grouped with the closely related RUBCN from the Penaeid species with more than 95 % identity. It was highly expressed in nerve followed by intestine, and gill. Its expression was induced upon WSSV challenge at 12 and 48 hpi. Suppression of PvRUBCN by dsRNA upon WSSV challenge resulted in more than 80 % reduction in PvRUBCN mRNA expression. Knockdown of PvRUBCN mRNA significantly decreased the WSSV copy number and prolonged the survival rate of WSSV-infected shrimp. In addition, the caspase3, a key regulator of the apoptosis pathway was significantly down-regulated. The interferon like genes including Vago4 was dramatically decreased at 72 hpi whereas Vago5 demonstrated slight reduction at 24 hpi and increase at 72 and 120 hpi, respectively. On the other hand, the expressions of the genes involved in the prophenoloxidase pathway including PPO1 and PPO2 were not changed. Taken together, PvRUBCN played important roles in the antiviral immunity in response to WSSV infection.