Molecular cloning, characterization and expression analysis of two juvenile hormone esterase-like carboxylesterase cDNAs in Chinese mitten crab, Eriocheir sinensis

Comp Biochem Physiol B Biochem Mol Biol. 2017 Mar:205:46-53. doi: 10.1016/j.cbpb.2017.01.002. Epub 2017 Jan 8.

Abstract

Precise regulation of methyl farnesoate (MF) titer is of prime importance throughout the crustacean life-cycle. Although the synthetic pathway of MF is well-documented, little is known about its degradation and recycling in crustaceans. Juvenile hormone esterase-like (JHE-like) carboxylesterase (CXE) is a key enzyme in MF degradation, thus playing a significant role in regulating the MF titer. We identified and characterized two cDNAs, Es-CXE1 and Es-CXE2, encoding JHE-like CXEs in Chinese mitten crab. Full-length cDNAs of Es-CXE1 and Es-CXE2 encode proteins composed of 584 and 597 amino acids, respectively, both of which contain a typical carboxylesterase domain. Alignment and phylogenetic analyses revealed that the Es-CXEs are highly similar to those of other crustaceans. To further validate their functions, we evaluated the mRNA expression patterns of the Es-CXEs in various tissues and in different physiological conditions. Tissue-specific expression analysis showed that the two Es-CXEs were predominantly expressed in the hepatopancreas and ovaries, which are the major tissues for MF metabolism. Es-CXE2 expression levels in the hepatopancreas and ovaries were about 100 and 25-fold higher, than the respective Es-CXE1 expressions. During ovarian rapid development stage, the global expressions of Es-CXEs were up-regulated in the hepatopancreas and down-regulated in the ovaries. After eyestalk ablation (ESA), the mRNA expressions of the two Es-CXEs were up-regulated in the hepatopancreas, further indicating their potential in degrading MF. Taken together, our results suggest that Es-CXEs, the key component of the juvenile hormone degradation pathway, may play vital roles in the development and reproduction of the Chinese mitten crab.

Keywords: Chinese mitten crab; Juvenile hormone esterase; Methyl farnesoate.

MeSH terms

  • Ablation Techniques
  • Amino Acid Sequence
  • Animals
  • Brachyura / enzymology*
  • Brachyura / genetics*
  • Brachyura / growth & development
  • Carboxylic Ester Hydrolases / chemistry
  • Carboxylic Ester Hydrolases / genetics*
  • Carboxylic Ester Hydrolases / metabolism*
  • Cloning, Molecular
  • DNA, Complementary / genetics*
  • Female
  • Gene Expression Regulation, Enzymologic*
  • Ovary / growth & development
  • Phylogeny
  • Protein Transport
  • Sequence Alignment
  • Structure-Activity Relationship

Substances

  • DNA, Complementary
  • Carboxylic Ester Hydrolases
  • juvenile hormone esterase