Cloning and characterization of a novel caspase-8-like gene in Crassostrea gigas

Fish Shellfish Immunol. 2015 Oct;46(2):486-92. doi: 10.1016/j.fsi.2015.06.035. Epub 2015 Jul 2.

Abstract

Cysteine-dependent aspartate-directed proteases, or caspases, play key roles in apoptosis and immune defense. In this study, we cloned the first caspase-8-like gene (CgCaspase8-2) identified in the pacific oyster, Crassostrea gigas. The 2572-bp cDNA encodes a putative protein of 714 amino acids that contains two tandem death effector domains (DEDs) at the N-terminal, and P20 and P10 domains at the C-terminal. The conserved pentapeptide motif QACQG was also identified in the deduced CgCaspase8-2 protein. Phylogenetic analysis indicated that CgCaspase8-2 was clustered with initiator caspases in the invertebrate subgroup, but the similarity between CgCaspase8-2 and other invertebrate caspase-8s was low. CgCaspase8-2 protein was localized in the cytoplasm, and over-expression of CgCaspase8-2 in HEK293T cells induced cell death, suggesting a role in apoptosis. Quantitative real-time PCR results demonstrated that CgCaspase8-2 was widely expressed in various tissues and developmental stages, with the highest CgCaspase8-2 expression levels detected in hemolymph and the blastula stage. Furthermore, CgCaspase8-2 transcripts showed no change in response to a bacterial challenge but exhibited notable up-regulation post-poly (I:C) challenge, suggesting that CgCaspase8-2 is specifically involved in immune responses against viruses. In summary, CgCaspase8-2 is involved in both apoptotic and immune function.

Keywords: Apoptosis; CgCaspase8-2; Crassostrea gigas; Immune response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Caspase 8 / genetics*
  • Caspase 8 / immunology
  • Caspase 8 / metabolism
  • Cell Survival
  • Cloning, Molecular
  • Crassostrea / genetics*
  • Crassostrea / immunology
  • Crassostrea / metabolism
  • DNA, Complementary / genetics
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Molecular Sequence Data
  • Phylogeny
  • Poly I-C / pharmacology
  • RNA, Messenger / metabolism
  • Sequence Analysis, DNA
  • Vibrio Infections / genetics
  • Vibrio Infections / immunology
  • Vibrio alginolyticus

Substances

  • DNA, Complementary
  • RNA, Messenger
  • Caspase 8
  • Poly I-C

Associated data

  • GENBANK/KR018807