Molecular cloning and characterization of an inhibitor of apoptosis protein in the mud crab Scylla paramamosain: its potential anti-apoptosis role under air exposure stress

The mud crab Scylla paramamosain is a widely farmed commercial species in southeast coastal areas of China. The crabs are placed in water-free containers for transportation to inland markets, thus are exposed to dry conditions for more than 72 h after capture and may suffer from air exposure stress, triggering cell apoptosis leading to death. To evaluate whether an inhibitor of apoptosis protein (IAP) is involved in apoptosis resistance against air exposure stress in crustaceans, SpIAP was cloned and investigated for the first time. The full length of SpIAP was 3351 bp, encoding a polypeptide of 662 amino acids. The predicted SpIAP protein contained three baculoviral IAP repeat domains and one really interesting new gene (RING) finger domain. Protein basic local alignment search tool and phylogenetic analysis results showed that SpIAP was clustered together with other crustaceans IAPs. SpIAP was detected in all the examined tissues and predominantly expressed in the hepatopancreas. When crabs were challenged with air exposure for 12 h, the expression level of SpIAP in the hepatopancreas was significantly increased in the experimental group compared with the control group. A RNA interference assay and flow cytometry analysis showed that when SpIAP was silenced, the cell apoptotic rate significantly increased after 24 h air exposure. These results suggested that SpIAP was involved in an anti-apoptosis response induced by air exposure in S. paramamosain.

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