Cloning and characterization of two ferritin subunit genes from bay scallop, Argopecten irradians (Lamarck 1819)

Mol Biol Rep. 2011 Mar;38(3):2125-32. doi: 10.1007/s11033-010-0339-0. Epub 2010 Sep 28.

Abstract

We have cloned two full-length cDNAs from two ferritin genes (Aifer1 and Aifer2) of the bay scallop, Argopecten irradians (Lamarck 1819). The cDNAs are 1,019 and 827 bp in length and encode proteins of 171 and 173 amino acids, respectively. The 5' UTR of each contains a conserved iron response element (IRE) motif. Sequence analyses reveal that both proteins belong to the H-ferritin family with seven conserved amino acids in the ferroxidase center. Highest expression of Aifer1 is found in the mantle and adductor muscle, while that of Aifer2 is only in the latter tissue. These Aifer genes are differentially expressed following bacterial challenge of the scallop. The expression level of Aifer1 was acutely up-regulated (over 10 fold) at 6 h post-bacteria injection, whereas Aifer2 expression was not significantly changed by bacterial challenge. Both genes were effectively expressed in E. coli BL21 (DE3), producing proteins of similar molecular weight, approximately 23 kDa. Purified Aifer1 and Aifer2 proteins exhibited iron-chelating activity of 33.1% and 30.4%, respectively, at a concentration of 5 mg/ml. Cations, Mg(2+), Zn(2+) and Ca(2+), depressed iron-chelating activity of both proteins. Additionally, the E. coli cells expressing recombinant Aifer1 and Aifer2 showed tolerance to H(2)O(2), providing a direct evidence of the antioxidation function of ferritin. The results presented in this study suggest important roles of Aifer1 and Aifer2 in the regulation of iron homeostasis, immune response, and antioxidative stress in A. irradians.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Biological Assay
  • Cloning, Molecular
  • Computational Biology
  • DNA, Complementary / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / drug effects
  • Escherichia coli / metabolism
  • Ferritins / genetics*
  • Ferritins / isolation & purification
  • Ferritins / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation / drug effects
  • Hydrogen Peroxide / pharmacology
  • Molecular Sequence Data
  • Pectinidae / genetics*
  • Pectinidae / microbiology
  • Protein Subunits / genetics*
  • Protein Subunits / isolation & purification
  • Protein Subunits / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Recombinant Proteins / metabolism
  • Sequence Analysis, DNA
  • Time Factors
  • Vibrio alginolyticus / drug effects
  • Vibrio alginolyticus / physiology

Substances

  • DNA, Complementary
  • Protein Subunits
  • RNA, Messenger
  • Recombinant Proteins
  • Ferritins
  • Hydrogen Peroxide

Associated data

  • GENBANK/GU263463
  • GENBANK/GU263464