title: Expression and functional characterization of a gene associated with retinoid-interferon-induced mortality 19 (GRIM-19) from orange-spotted grouper (Epinephelus coioides).
abstract: GRIM-19 is a nuclear encoded subunit of complex I that has been implicated in apoptosis. The protein participates in multiple functions including the innate immune response. GRIM-19 has been studied in humans and other mammals; however, fish GRIM-19 has not been well characterized. In this study, a new GRIM-19 gene, EcGRIM-19, was isolated from the orange-spotted grouper (Epinephelus coioides) cDNA library, which was constructed following LPS treatment. EcGRIM-19 is a 582-bp gene that encodes a 144-amino acid protein. The gene is a true ortholog of mammalian GRIM-19. EcGRIM-19 exhibits ubiquitous and constitutive expression in the different tissues of the orange-spotted grouper. The expression levels of EcGRIM-19 are altered in the gill, spleen, kidney and liver after induction with LPS. The subcellular localization analysis demonstrated that the EcGRIM-19 protein is localized predominantly in the mitochondria. In addition, amino acids 30-50 of the protein are responsible for the mitochondrial localization of EcGRIM-19. The caspase assay demonstrated that the overexpression of GRIM-19 enhanced the cellular sensitivity to interferon(IFN)-β- and retinoic acid (RA)-induced death in HeLa cells. The data presented in this study are important for further understanding the EcGRIM-19 gene function in fish.
authors.affiliation: Key Laboratory of Tropical & Subtropical Fishery Resource Application & Cultivation, Ministry of Agriculture, Pearl River Fishery Research Institute, Chinese Academic of Fishery Science, Guangzhou 510380, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Shi
authors.forename: Zhe
authors.initials: Z
authors.lastname: Zhao
authors.forename: Xinping
authors.initials: X
authors.lastname: Zhu
authors.forename: Kunci
authors.initials: K
authors.lastname: Chen
authors.forename: Qiya
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D001492:Bass / Q000235:genetics* / Q000378:metabolism*; D020169:Caspases / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D016899:Interferon-beta / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010802:Phylogeny; D012313:RNA / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment; D014162:Transfection; D014212:Tretinoin / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D029941:Fish Proteins; D008070:Lipopolysaccharides; D014212:Tretinoin; D012313:RNA; D016899:Interferon-beta; D020169:Caspases