PUBLICATION

Cloning and functional characterization of the p65 subunit of NF-κB from olive flounder (Paralichthys olivaceus)

Authors
Kong, H.J., Moon, J.H., Moon, J.Y., Kim, J.M., Nam, B.H., Kim, Y.O., Kim, W.J., and Lee, S.J.
ID
ZDB-PUB-101209-23
Date
2011
Source
Fish & shellfish immunology   30(1): 406-411 (Journal)
Registered Authors
Kong, Hee Jeong
Keywords
p65, NF-κB, Paralichthys olivaceus, Transactivation, Poly I:C
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Cloning, Molecular
  • Flounder*
  • Gene Expression Regulation/physiology*
  • Molecular Sequence Data
  • NF-kappa B/genetics
  • NF-kappa B/metabolism*
  • Protein Subunits
  • Protein Transport
  • RNA, Messenger/genetics
  • RNA, Messenger/metabolism
PubMed
21130886 Full text @ Fish Shellfish Immunol.
Abstract
NF-κB is a master transcription factor found in almost all cell types that responds to diverse cellular stimuli by activating the expression of stress response genes, including immune-related genes. cDNA encoding the p65 subunit of olive flounder (Paralichthys olivaceus) NF-κB (Po-p65) was isolated through an EST analysis of an olive flounder cDNA library, a screen of BAC library, and rapid amplification of cDNA ends (RACE). The cDNA for Po-p65 encodes a polypeptide 626 amino acids in length containing a well-conserved Rel-homology domain (RHD). The primary sequence of Po-p65 showed strong homology with p65 from perch and zebrafish (82.7 and 64.4%, respectively), and shared 43.4-42.1% homology with p65 from other species, including mammals, while the N-terminal RHD of Po-p65 showed strong identity (95.6-67.8%) with that of other species. Po-p65 mRNA expression was detected in all flounder tissues examined. The over-expression of full-length Po-p65 (Po-p65f), but not of a Po-p65 C-terminus deletion mutant (Po-p65ΔC), stimulated κB element-driven reporter (κB-luc) activity in a dose-dependent manner and regulated the expression of p65 target genes, including TNF-α and IκB-α,, in HINAE olive flounder cells. Po-p65f translocated to the nucleus following stimulation with poly I:C in HINAE cells. Together, these results suggest that Po-p65 is evolutionarily and functionally conserved in flounder and mammals and may provide clues to the detailed molecular mechanism(s) underlying immune response regulation in flounder.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping