H2A and Ca-L-hipposin gene: Characteristic analysis and expression responses to Aeromonas hydrophila infection in Carassius aurutus.

Antimicrobial peptide is an important component of the host innate immune system and thus serves a crucial function in host defense against microbial invasion. In this study, H2A and derived antimicrobial peptide Ca-L-hipposin were cloned and characterized in Carassius aurutus. The gene H2A full-length cDNA is 908 bp and includes a 5'-terminal untranslated region (UTR) of 55 bp and a 3'-terminal UTR of 466 bp with a canonical polyadenylation signal sequence AATAA, as well as an open reading frame (ORF) of 387 bp encoding a polypeptide of 128 amino acids, with a molecular weight of 13.7 kDa, an isoelectric point of 10.7, and 94% homology with Danio rerio H2A. The secondary structure of H2A includes the α-spiral with 51 amino acids with a composition ratio of 39.8%, as well as a β-corner with 15 amino acids in a composition ratio of 11.7%. The online software ExPaSy predicted that a peptide sequence with 51 amino acids from the 2nd to 52nd amino acids in histone H2A can be produced through hydrolization by protease chymotrypsin, which indicates a difference of only three amino acids, compared with the antimicrobial peptide hipposin in Hippoglossus hippoglossus with a homology of 94%. Ca-L-hipposin includes 51 amino acids with a molecular weight of 5.4 kDa and an isoelectric point of 12.0, the secondary structure of which contains an α-helix of 17 amino acids accounting for 33.3% and a β-corner of 8 amino acids accounting for 15.7%. H2A was extensively expressed in the mRNA levels of various tissues, with higher expression levels in kidney and spleen. After C. aurutus was challenged with Aeromonas hydrophila, the mRNA expression levels of H2A were upregulated in the kidney, spleen, and liver. H2A serves an important function in the defense against the invasion of A. hydrophila. In addition, sequence characteristics reveal that Ca-L-hipposin could be a potential antimicrobial peptide for use in killing pathogenic bacteria in aquaculture.