Molecular cloning, characterization and expression analysis of cathepsin O in silkworm Bombyx mori related to bacterial response

Mol Immunol. 2015 Aug;66(2):409-17. doi: 10.1016/j.molimm.2015.04.008. Epub 2015 May 18.

Abstract

Cathepsins are the main members of the cysteine family and play important roles in immune response in vertebrates. The Cathepsin O of Bombyx mori (BmCathepsin O) was cloned from the hemocytes by the rapid amplification of cDNA ends (RACE). The genomic DNA was 6131bp long with a total of six exons and five introns. Its pre-mRNA was spliced to generate two spliceosomes. By comparisons with other reported cathepsins O, it was concluded that the identity between them ranged from 29 to 39%. Expression analysis indicated that BmCathepsin O was specific-expressed in hemocytes, and highly expressed at the 4th molting and metamorphosis stages. Immunofluorescence assay and qRT-PCR showed that BmCathepsin O was expressed in granulocytes and plasmatocytes. Interestingly, BmCathepsin O was significantly up-regulated after stimulated by 20-hydroxyecdysone (20-E) in vivo, which suggested that BmCathepsin O may be regulated by 20E. Moreover, activation of BmCathepsin O was also observed in hemocytes challenged by Escherichia coli, indicating its potential involvement in the innate immune system of silkworm, B. mori. In summary, our studies provide a new insight into the functional features of Cathepsin O.

Keywords: 20-Ecdysone; Cathepsin O; Infection; Molecular cloning; Silkworm.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Bombyx / classification
  • Bombyx / drug effects
  • Bombyx / genetics
  • Bombyx / immunology*
  • Cathepsins / genetics
  • Cathepsins / immunology*
  • Cloning, Molecular
  • Ecdysterone / pharmacology
  • Escherichia coli / immunology
  • Exons
  • Gene Expression
  • Hemocytes / immunology*
  • Hemocytes / microbiology
  • Immunity, Innate*
  • Insect Proteins / genetics
  • Insect Proteins / immunology*
  • Introns
  • Metamorphosis, Biological / immunology
  • Molecular Sequence Data
  • Open Reading Frames
  • Phylogeny
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Sequence Alignment
  • Sequence Homology, Amino Acid

Substances

  • Insect Proteins
  • Recombinant Proteins
  • Ecdysterone
  • Cathepsins