Molecular identification and functional analysis of MyD88 in giant freshwater prawn (Macrobrachium rosenbergii) and expression changes in response to bacterial challenge

Int J Biol Macromol. 2021 May 1:178:492-503. doi: 10.1016/j.ijbiomac.2021.02.177. Epub 2021 Feb 26.

Abstract

Myeloid differentiation factor 88 (MyD88) is a crucial adaptor protein for Toll-like receptor (TLR)-mediated signaling pathways and plays an important role in immune response. In this study, the full-length cDNA of MyD88 from Macrobrachium rosenbergii (MRMyD88) was cloned. The MRMyD88 cDNA is 1758 bp long and contains a 1398-bp open reading frame. Multiple sequence alignment and phylogenetic analysis revealed that the amino acid sequence of MRMyD88 shared high identity with the known MyD88 proteins. The MRMyD88 mRNA was widely expressed in all examined tissues, with highest level in intestine, followed by gonad and pleopod. Furthermore, the MRMyD88 promoter region, spanning 1622 bp, contains several transcription factor-binding sites, including nine GATA-1 box motifs. Electrophoretic mobility shift assay showed that Gfi-1, SRF, and Oct-1 bind to the upstream region of MRMyD88. Additionally, the results showed that the expression levels of TLR1, TLR2 and TLR3 were different in response to Vibrio anguillarum, Lactobacillus plantarum and Aeromonas hydrophila infections. However, these bacteria significantly increased the expression levels of MyD88 and prophenoloxidase. These data suggest that the TLR-mediated signaling pathway is MyD88-dependent in response to pathogenic and probiotic bacteria in M. rosenbergii.

Keywords: Cloning; Macrobrachium rosenbergii; MyD88.

MeSH terms

  • Animals
  • Arthropod Proteins* / genetics
  • Arthropod Proteins* / immunology
  • Myeloid Differentiation Factor 88* / genetics
  • Myeloid Differentiation Factor 88* / immunology
  • Palaemonidae* / genetics
  • Palaemonidae* / immunology
  • Palaemonidae* / microbiology
  • Vibrio / immunology*
  • Vibrio Infections* / genetics
  • Vibrio Infections* / immunology

Substances

  • Arthropod Proteins
  • Myeloid Differentiation Factor 88

Supplementary concepts

  • Vibrio anguillarum