Minichromosome maintenance protein 7 regulates phagocytosis in kuruma shrimp Marsupenaeus japonicas against white spot syndrome virus.
Minichromosome maintenance protein (MCM7) belongs to the MCM protein family and participates in the MCM complex by playing a role in the cell replication cycle and chromosome initiation of eukaryotes. Previously, we found that several genes, including MCM7, were over-expressed in Drosophila melanogaster after white spot syndrome virus (WSSV) infection. In this study, we aimed to further research the MCM7 of kuruma shrimp, Marsupenaeus japonicus (mjMCM7) and determine its role in the innate immune system. To this end, we cloned the entire 2307-bp mjMCM7 sequence, including a 1974-bp open reading frame (ORF) encoding a 658-aa-long protein. Real-time PCR showed that the gene was primarily expressed in the hemolymph and hepatopancreas and over-expressed in shrimp challenged with WSSV. Gene function study was carried out by knocking down the expression of MCM7 using small interference RNA (siRNA). The results revealed that β-actin, hemocyanin, prophenoloxidase (proPO) and tumor necrosis factor-α (TNF-α) were up-regulated while the cytoskeleton proteins such as myosin and Rho were significantly down-regulated at 24 h after treatment. The results indicate a possible relationship between mjMCM7 and the innate immune system, and suggest that mjMCM7 may play a role in phagocytosis. After WSSV challenge, WSSV copies and mortality count were both higher in the MCM7-siRNA-treated groups at 60 h after treatment, and the mortality count approached that of the control groups over time. The phagocytosis rate was significantly lower in the MCM7-siRNA-treated group than in the WSSV group. The findings of this study confirm that mjMCM7 positively regulates phagocytosis and plays an important role against WSSV. These results could help researchers to further understand the function of the MCM7 protein and reveal its potential role in the innate immunity of invertebrates.
College of Animal Science and Technology, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China.
Zhi
Z
Wang
College of Animal Science and Technology, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China. Electronic address: zhufei@zju.edu.cn.
Fei
F
Zhu
D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D007113:Immunity, Innate / Q000235:genetics*; D064168:Minichromosome Maintenance Complex Component 7 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D033561:Penaeidae / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism / Q000821:virology; D010587:Phagocytosis; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment; D046848:White spot syndrome virus 1 / Q000502:physiology*
D060829:Arthropod Proteins; D018076:DNA, Complementary; D064168:Minichromosome Maintenance Complex Component 7
Gene expression; Marsupenaeus japonicus; Minichromosome maintenance protein 7; Phagocytosis; White spot syndrome virus