A novel nuclear factor Akirin regulating the expression of antimicrobial peptides in Chinese mitten crab Eriocheir sinensis.

Akirin, a recently discovered nuclear factor, participates in regulating various processes, including cell proliferation and differentiation, embryonic development, and immunity. In the present study, a novel Akirin was identified from Chinese mitten crab Eriocheir sinensis (designated as EsAkirin), and its primary functions in regulating antimicrobial peptides were explored. The open reading frame of EsAkirin was of 615 bp, encoding a polypeptide of 204 amino acid residues. The deduced amino acid sequence of EsAkirin shared high similarities ranging from 44.1% to 89.2% with other Akirins. In the phylogenetic tree, EsAkirin was firstly clustered with Akirins from shrimp and then assigned into the invertebrate branch. The mRNA transcripts of EsAkirin were constitutively expressed in all the tested tissues, with the highest expression level (5.07-fold compared to the stomach, p < 0.01) in hepatopancreas. The mRNA expression of EsAkirin in hemocytes was significantly increased at 6 h, and reached the maximum level at 24 h post stimulations with either lipopolysaccharide (LPS) (5.04-fold, p < 0.01) or Aeromonas hydrophila (3.10-fold, p < 0.01). After the injection of EsAkirin-dsRNA, the mRNA expressions of EsALF2, EsLYZ, EsCrus2 and EsDWD1 were significantly decreased (p < 0.01) upon LPS stimulation. EsAkirin protein was prominently distributed in the nucleus of E. sinensis hemocytes after LPS and A. hydrophila stimulations. The relative luciferase reporter system analysis revealed that the activity of nuclear factor-κB was significantly up-regulated (2.64-fold, p < 0.01) in human embryonic kidney (HEK293T) cells after the over-expression of EsAkirin. Collectively, these results suggested that EsAkirin might play an important role in the immune responses of E. sinensis by regulating the expression of antimicrobial peptides.