title: Molecular characterization and gene expression of ferritin in blunt snout bream (Megalobrama amblycephala).
abstract: Ferritins are conserved iron storage proteins that exist in most living organisms and play an essential role in iron homeostasis. In this study, we reported the identification and analysis of a ferritin middle-chain (M) subunit, MaFerM, from blunt snout bream, Megalobrama amblycephala. The full length cDNA of MaFerM contains a 5'-untranslated region (UTR) of 152 bp, an open reading frame (ORF) of 522 bp and a 3'-UTR of 270 bp. The ORF encodes a putative protein of 174 amino acids, which shares extensive sequence identities with the M ferritins of several fish species. In silico analysis identified both the ferroxidase center of mammalian heavy-chain (H) ferritins and the iron nucleation site of mammalian light-chain (L) ferritins in MaFerM. Quantitative real-time reverse transcription polymerase chain reaction analysis indicated that MaFerM expression was highest in the liver and lowest in the heart and responded positively to experimental challenges with Aeromonas hydrophila. The exposure of cultured M. amblycephala to treatment with stress inducers (iron and H2O2) significantly up-regulated the expression of MaFerM in a dose-dependent manner. Iron chelation analysis showed that recombinant MaFerM purified from Escherichia coli exhibited apparent iron binding activity. These results suggest that MaFerM is a functional M ferritin and is likely to play a role in iron sequestration and protection against oxidative stress and immune stimulus.
authors.affiliation: Key Laboratory of Genetic Breeding and Aquaculture Biology of Freshwater Fishes, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, No. 9 East Shanshui Road, Wuxi, Jiangsu 214081, PR China.
authors.forename: Shengming
authors.initials: S
authors.lastname: Sun
authors.affiliation: Key Laboratory of Genetic Breeding and Aquaculture Biology of Freshwater Fishes, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, No. 9 East Shanshui Road, Wuxi, Jiangsu 214081, PR China. Electronic address: zhuj@ffrc.cn.
authors.forename: Jian
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Genetic Breeding and Aquaculture Biology of Freshwater Fishes, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, No. 9 East Shanshui Road, Wuxi, Jiangsu 214081, PR China. Electronic address: gexp@ffrc.cn.
authors.forename: Xianping
authors.initials: X
authors.lastname: Ge
authors.affiliation: Wuxi Fishery College Nanjing Agricultural University, Wuxi 214081, PR China.
authors.forename: Wxuxiao
authors.initials: W
authors.lastname: Zhang
mesh_terms: D016980:Aeromonas hydrophila / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003530:Cyprinidae; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005293:Ferritins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005393:Fish Diseases / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D016905:Gram-Negative Bacterial Infections / Q000235:genetics / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D007113:Immunity, Innate; D018384:Oxidative Stress; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment / Q000662:veterinary; D014018:Tissue Distribution
chemical_list: D018076:DNA, Complementary; D029941:Fish Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D005293:Ferritins
keywords: Bacterial infection; Ferritin; Iron; Megalobrama amblycephala; Oxidative stress