PUBLICATION
Molecular cloning and characterization of two types of IkappaBalpha orthologues in orange-spotted grouper, Epinephelus coioides
- Authors
- Gao, R., Huang, Y., Huang, X., Guan, L., Wei, S., Zhou, Y., Qin, Q.
- ID
- ZDB-PUB-140513-419
- Date
- 2014
- Source
- Fish & shellfish immunology 38: 101-10 (Journal)
- Registered Authors
- Keywords
- EcIκBαA, EcIκBαB, Epinephelus coioides, NF-κB, SGIV
- MeSH Terms
-
- Tissue Distribution
- Perciformes/metabolism*
- Transcriptome
- Base Sequence
- Protein Transport
- Molecular Sequence Data
- I-kappa B Proteins/genetics
- I-kappa B Proteins/metabolism*
- Spleen/cytology
- Animals
- Amino Acid Sequence
- Phylogeny
- Cloning, Molecular
- Cell Line
- Gene Expression Regulation/physiology
- PubMed
- 24594011 Full text @ Fish Shellfish Immunol.
Citation
Gao, R., Huang, Y., Huang, X., Guan, L., Wei, S., Zhou, Y., Qin, Q. (2014) Molecular cloning and characterization of two types of IkappaBalpha orthologues in orange-spotted grouper, Epinephelus coioides. Fish & shellfish immunology. 38:101-10.
Abstract
Inhibitors of kappa B (LκBs) are the members of primary regulators of NF-κB, which can inhibit NF-κB activity by blocking the NF-κB in an inactive state in the cytoplasm. In this study, two types of LκBα orthologues (EcLκBαA and EcLκBαB) from orange-spotted grouper, Epinephelus coioides, were cloned and characterized. EcLκBαA and EcLκBαB encoded putative proteins containing 308 and 318 amino acids, which shared 59% and 53% identity to LκBαA and LκBαB of Danio rerio, respectively. Amino acid sequence alignment showed that both EcLκBαA and EcLκBαB contained a conserved degradation motif DSGLDS in the N-terminal region and a PEST sequence in the C-terminal region. In addition, EcLκBαA and EcLκBαB contained 5 and 6 ankyrin repeats, respectively. The genomic DNA of EcLκBαA and EcLκBαB consisted of 6 exons and 5 introns. Both of their transcripts were widely distributed in different tissues, and the expression levels were different in response to various stimuli, including lipopolysaccharide (LPS), Vibrio alginolyticus and Singapore grouper iridovirus (SGIV). Dual-luciferase reporter assay suggested that both EcLκBαA and EcLκBαB were able to inhibit Ecc-Rel and Ecp65 induced NF-κB promoter activity in grouper spleen (GS) cells. Subcellular localization analysis showed that EcLκBαB was present predominantly in the cytoplasm, while EcLκBαA was distributed throughout both the nucleus and the cytoplasm. Furthermore, overexpression of EcLκBαA and EcLκBαB in GS cells inhibited the viral gene transcriptions of MCP, ORF019 and ORF162 of SGIV. Taken together, our findings suggested that both EcLκBαA and EcLκBαB were involved in grouper innate immunity against virus.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping