Identification and expression of a putative LPS-induced TNF-α factor from Asiatic hard clam Meretrix meretrix

Mol Biol Rep. 2012 Feb;39(2):865-71. doi: 10.1007/s11033-011-0810-6. Epub 2011 May 13.

Abstract

LPS-induced TNF-α (LITAF) is a novel transcriptional factor that mediates the expression of inflammatory cytokines in LPS-induced processes. In the present study, the full-length cDNA encoding LITAF (designated as Mm-LITAF) was identified from Asiatic hard clam, Meretrix meretrix, by expressed sequence tag and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of Mm-LITAF was 1653 bp, consisting of a 5' untranslated region (UTR) of 91 bp, a 3'UTR of 1166 bp with one cytokine RNA instability motif (ATTTA) and one polyadenylation signal (AATAAA), and an open reading frame (ORF) of 396 bp encoding a polypeptide of 131 amino acids with a theoretical isoelectric point of 7.49, and predicted molecular weight of 14.47 kDa. The deduced amino acid of Mm-LITAF shared 29-63% similarity with the LITAFs from other species, indicating that Mm-LITAF should be a member of the LITAF family. Two highly conserved CXXC motifs forming a compact Zn(2+)-binding structure were also identified in Mm-LITAF. A quantitative reverse transcriptase real-time PCR (qRT-PCR) assay was developed to assess the expression of Mm-LITAF mRNA in different tissues, and the temporal expression of Mm-LITAF in clams challenged with Vibrio anguillarum. The mRNA transcript of Mm-LITAF could be detected in all the examined tissues with the highest expression level in the gill. Mm-LITAF expression was up-regulated significantly at 16 h in the gill and at 8 h in haemocytes after bacterial challenge, respectively. These results suggest that the Mm-LITAF is a constitutive and inducible acute-phase protein that perhaps involved in the innate immune response of hard clam.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Bivalvia / genetics*
  • Bivalvia / microbiology
  • Cloning, Molecular
  • Cluster Analysis
  • DNA Primers / genetics
  • DNA, Complementary / genetics
  • Expressed Sequence Tags
  • Gene Components
  • Gills / metabolism
  • Hemocytes / metabolism
  • Lipopolysaccharides
  • Molecular Sequence Data
  • Open Reading Frames / genetics
  • Phylogeny*
  • RNA, Messenger / genetics*
  • Real-Time Polymerase Chain Reaction
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Tumor Necrosis Factor-alpha / genetics*
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • DNA Primers
  • DNA, Complementary
  • Lipopolysaccharides
  • RNA, Messenger
  • Transcription Factors
  • Tumor Necrosis Factor-alpha