Identification and characterization of nascent polypeptide-associated complex alpha from Chinese mitten crab (Eriocheir sinensis): A novel stress and immune response gene in crustaceans

• Eriocheir sinensis nascent polypeptide-associated complex alpha gene (EsNACA) was cloned.
• EsNACA was constitutively expressed with dominant expression in hepatopancreas.
• EsNACA gene expression was significantly altered by salinity and pH stresses.
• EsNACA gene expression was significantly up-regulated by PAMPs and bacterial challenges.
• EsNACA is an inducible gene involved in stress and immune response in crustaceans.

Disease in aquatic animals is tightly linked to environmental challenges and their immune responses are greatly modified by their external environment. The chaperone protein nascent polypeptide-associated complex alpha (NACA) has been suggested to play important roles in the cellular response to stress and immune challenges, while the related biological functions remain largely unknown in invertebrates. In the present study we identified a NACA gene (termed EsNACA) from Chinese mitten crab Eriocheir sinensis and analyzed its expression changes in response to ambient (salinity and pH) stresses and immune challenges. The EsNACA protein is comprised of 209 amino acid residues with a conserved DNA binding domain, a C-Jun binding domain, a NAC domain and an ubiquitin-associated domain and shows the highest sequence identity (87%) with its counterpart in shrimp Penaeus monodon. EsNACA mRNA transcripts are presented in all tested normal tissues with predominant expression in hepatopancreas and lower expression in hemocytes. In addition, EsNACA expression was significantly altered in response to the ambient salinity (15‰ and 30‰ salinities) and pH (pH 6 and 8.5) stresses in gill, hepatopancreas, muscle, hemocytes and intestine tissues. Furthermore, EsNACA gene expression was substantially induced upon LPS and Poly(I:C) immune stimulations in E. sinensis hemocytes in vitro. Finally, EsNACA expression was up-regulated in E. sinensis hemocytes, gill, hepatopancreas, intestine and muscle tissues in response to Vibrio anguillarum challenges in vivo. Taken together, our findings for the first time show that EsNACA is an inducible gene involved in stress and immune response in crustaceans.