An interaction between nervous necrosis virus capsid protein and nucleolar phosphoprotein B23 was identified. The capsid protein binds to B23 early during the viral infection and accumulates in the nucleolus. Knockdown of B23 reduces the cytopathic effect (CPE) and replication efficiency of NVV. The interaction of B23 with NNV capsid protein may represent an important mechanism in NNV replication.