Molecular characterization of minichromosome maintenance protein (MCM7) in Scylla paramamosain and its role in white spot syndrome virus and Vibrio alginolyticus infection.

The minichromosome maintenance protein (MCM7) is a member of the MCM protein family which participates in the MCM complex by playing a role in the cell replication cycle and chromosome initiation in eukaryotes. The 2270 bp cDNA sequence of MCM7, including a 2127-bp open reading frame (ORF) encoding a 709-aa protein, was cloned from Scylla paramamosain using RT-PCR and RACE. Data showed that MCM7 was highly expressed in the digestive organ and hepatopancreas of S. paramamosain. Furthermore, MCM7 expression was down-regulated by infection with white spot syndrome virus (WSSV) or Vibrio alginolyticus. When MCM7 was knocked down, immune genes such as Janus kinase (JAK) and crustin antimicrobial peptide (CAP) were down-regulated, and C-type-lectin (CTL) was up-regulated in hemocytes. The mortality of WSSV-infected or V. alginolyticus-infected crabs was enhanced following MCM7 knockdown. It was demonstrated that MCM7 is very important in the progression of WSSV and V. alginolyticus infection. We also investigated the effect of MCM7 on apoptosis rate and phagocytic rate in S. paramamosain. MCM7 knockdown caused higher levels of apoptosis in the hemocytes of the control, WSSV, and V. alginolyticus groups. MCM7 knockdown influenced the activity of phenoloxidase (PO) and superoxide dismutase (SOD), and total hemocyte count (THC) after infection with WSSV or V. alginolyticus, which indicated that MCM7 plays a regulatory role in innate immunity of crabs. Thus, we conclude that MCM7 may participate in the anti-WSSV and V. alginolyticus immune response in crabs by regulating apoptosis and the activity of PO and SOD.