title: Molecular profile and functional characterization of the ferritin H subunit from rock bream (Oplegnathus fasciatus), revealing its putative role in host antioxidant and immune defense.
abstract: Ferritins are iron binding proteins made out of 24 subunits, involved in iron homeostasis and metabolism in cellular environments. Here, we sought to identify and functionally characterize a one type of subunits of ferritin (ferritin H-like subunit) from rock bream (Oplegnathus fasciatus; RbFerH). The complete coding sequence of RbFerH was 531 bp in length, encoding a 177-amino acid protein with a predicted molecular mass of 20.8 kDa. The deduced protein structure possessed the domain architecture characteristic of known ferritin H subunits, including metal ligands for iron binding, a ferroxidase center, and two iron-binding region signatures. As expected, the 5' untranslated region of the RbFerH cDNA sequence contained a putative iron response element region, a characteristic regulatory element involved in its translation. The RbFerH gene comprised 5 exons and 4 introns spanning a 4195 bp region. Overexpressed recombinant RbFerH protein demonstrated prominent Fe(II) ion depriving activity, bacteriostatic properties, and protective effects against oxidative double-stranded DNA damage. Using quantitative polymerase chain reaction (qPCR), we found that RbFerH was expressed ubiquitously in the majority of physiologically important tissues in rock bream. A greater abundance of the mRNA transcripts were detected in blood and liver tissues. Upon administering different microbial pathogens and pathogen-derived mitogens, RbFerH transcription was markedly elevated in the blood of rock bream. Taken together, our findings suggest that RbFerH acts as a potent iron sequestrator in rock bream and may actively participate in antimicrobial as well as antioxidative defense.
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea.
authors.forename: Don Anushka Sandaruwan
authors.initials: DA
authors.lastname: Elvitigala
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea.
authors.forename: Thanthrige Thiunuwan
authors.initials: TT
authors.lastname: Priyathilaka
authors.affiliation: Fish Vaccine Research Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea.
authors.forename: Bong-Soo
authors.initials: BS
authors.lastname: Lim
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea.
authors.forename: Ilson
authors.initials: I
authors.lastname: Whang
authors.affiliation: Department of Biotechnology, Division of Applied Chemistry & Biotechnology, Hanbat National University, Daejeon 305-719, Republic of Korea.
authors.forename: Sang-Yeob
authors.initials: SY
authors.lastname: Yeo
authors.affiliation: Division of Marine Environment and Bioscience, Korea Maritime University, Busan 606-791, Republic of Korea.
authors.forename: Cheol Young
authors.initials: CY
authors.lastname: Choi
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea. Electronic address: jehee@jejunu.ac.kr.
authors.forename: Jehee
authors.initials: J
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001052:Apoferritins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D004249:DNA Damage; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D007501:Iron / Q000378:metabolism; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010473:Perciformes; D010802:Phylogeny; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment
chemical_list: D029941:Fish Proteins; D011993:Recombinant Fusion Proteins; D001052:Apoferritins; D007501:Iron
keywords: Ferritin H; Functional analysis; Pathogen-induced expressional regulation; Recombinant protein; Rock bream