20-Hydroxyecdysone regulates the transcription of the lysozyme via Broad-Complex Z2 gene in silkworm, Bombyx mori

Dev Comp Immunol. 2019 May:94:66-72. doi: 10.1016/j.dci.2019.01.014. Epub 2019 Feb 1.

Abstract

Broad-Complex Z2 (Br-C Z2) is an ecdysone inducible transcription factor that regulates physiological, innate immune and developmental events in insects. Here, we identified an orthologue of Br-C Z2 from silkworm, Bombyx mori (BmBr-C Z2) to study its involvement in immune responses. The quantitative real-time PCR analysis revealed that BmBr-C Z2 was expressed ubiquitously in all tested tissues under normal physiological conditions. Further, developmental profile displayed that BmBr-C Z2 expression was detectable in different developmental stages, however the gene's expression was highest in the molting and pre-pupal stages. Administration of 20-hydroxyecdysone (20E) enhanced the expression levels of BmBr-C Z2 in hemocytes. The challenge with pathogens and pathogen associated molecular patterns (PAMPs) also upregulated the mRNA levels of BmBr-C Z2 in hemocytes when compared with the control. By contrast, the ectopic expression of BmBr-C Z2 remarkably increased the production of antimicrobial peptides, while the knock-down of this gene by double stranded RNA decreased their production. Dual-luciferase assay exhibited that BmBr-C Z2 induced the expression of lysozyme by directly binding to its promoter region. The treatment of Escherichia coli following the knock-down of BmBr-C Z2 strongly reduced the survival rate of silkworm larvae. These results suggest that BmBr-C Z2 plays an important biological role in the innate immune responses of silkworm by regulating immune-related genes.

Keywords: 20E; AMPs; BmBr-C Z2; Bombyx mori; Insect immunity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antimicrobial Cationic Peptides / genetics
  • Antimicrobial Cationic Peptides / metabolism
  • Bombyx / physiology*
  • Cloning, Molecular
  • Drosophila Proteins / genetics
  • Ecdysterone / metabolism*
  • Escherichia coli / physiology*
  • Escherichia coli Infections / immunology*
  • Gene Expression Regulation, Developmental
  • Immunity, Innate
  • Insect Proteins / genetics*
  • Insect Proteins / metabolism
  • Metamorphosis, Biological
  • Muramidase / genetics
  • Muramidase / metabolism
  • Promoter Regions, Genetic / genetics
  • RNA, Small Interfering / genetics
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Transcriptional Activation

Substances

  • Antimicrobial Cationic Peptides
  • Br protein, Drosophila
  • Drosophila Proteins
  • Insect Proteins
  • RNA, Small Interfering
  • Transcription Factors
  • Ecdysterone
  • Muramidase