title: A teleostean counterpart of ferritin M subunit from rock bream (Oplegnathus fasciatus): an active constituent in iron chelation and DNA protection against oxidative damage, with a modulated expression upon pathogen stress.
abstract: Ferritins are biological iron chelators that can sequestrate excess iron to maintain iron homeostasis in the body. Ferritins basically consist of 2 types of subunits, designated as H and L. However, another new subunit, ferritin "M" which possesses characteristic features of both the H and L subunits, was recently identified in lower vertebrates, mostly in fish. In this study, a ferritin M-like subunit from rock bream (Oplegnathus fasciatus) (RbFerM) was characterized at the molecular level, and its transcriptional profile was analyzed in healthy fish, as well as in pathogen- and mitogen-stimulated fish. Furthermore, its functional properties were evaluated using the recombinant protein. The complete coding sequence of RbFerM was 528 bp in length, encoding a 176-amino acid peptide with a calculated molecular mass of 20 kDa. In silico analysis of RbFerM revealed that it has features similar to both the mammalian ferritin subunits, H and L. Phylogenetic analysis depicted the higher evolutionary proximity of RbFerM with its fish counterparts. Quantitative real time polymerase chain reaction (PCR) analysis detected a ubiquitous transcriptional profile of RbFerM in selected tissues of rock bream, in which more pronounced expression was observed in blood and liver tissues. Significant transcriptional inductions of RbFerM were detected in liver tissues upon lipopolysaccharides (LPS), Edwardsiella tarda, Streptococcus iniae, and rock bream irido virus (RBIV) exposures in time-course immune-challenge experiments. The purified recombinant protein of RbFerM demonstrated detectable iron chelating activity that varied with the temperature. Moreover, the recombinant RbFerM rendered a detectable protection effect against iron (II) and H2O2-mediated DNA damage.
authors: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea.
authors: Don Anushka Sandaruwan
authors: DA
authors: Elvitigala
authors: H K A
authors: HK
authors: Premachandra
authors: Ilson
authors: I
authors: Whang
authors: Myung-Joo
authors: MJ
authors: Oh
authors: Sung-Ju
authors: SJ
authors: Jung
authors: Choul-Ji
authors: CJ
authors: Park
authors: Jehee
authors: J
authors: Lee
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D001483:Base Sequence; D016000:Cluster Analysis; D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D020609:Edwardsiella tarda / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D005293:Ferritins / Q000235:genetics* / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000662:veterinary; D017994:Iridovirus / Q000276:immunology; D007502:Iron Chelating Agents / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000276:immunology; D008969:Molecular Sequence Data; D010473:Perciformes / Q000235:genetics* / Q000276:immunology*; D010802:Phylogeny; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D011994:Recombinant Proteins / Q000378:metabolism*; D012394:Rosaniline Dyes; D016415:Sequence Alignment / Q000662:veterinary; D013291:Streptococcus / Q000276:immunology
chemical_list: D000975:Antioxidants; D017931:DNA Primers; D007502:Iron Chelating Agents; D008070:Lipopolysaccharides; D021122:Protein Subunits; D011994:Recombinant Proteins; D012394:Rosaniline Dyes; C048139:Coomassie blue; D005293:Ferritins
keywords: DNA protection effect; Ferritin M; Iron chelation; Rock bream; Transcriptional analysis