Interleukin-1beta gene in orange-spotted grouper, Epinephelus coioides: molecular cloning, expression, biological activities and signal transduction.
IL-1beta, a key mediator of inflammation, orchestrates a variety of immune responses by initiating gene expression. Herein, we have cloned and sequenced the IL-1beta in orange-spotted grouper (Epinephelus coioides), produced soluble mature recombinant IL-1beta in Escherichia coli, and characterized its biological properties and downstream signal transduction. The grouper IL-1beta cDNA was 1364bp in length, containing an open reading frame of 765bp. The predicted protein of 254 amino acids revealed the presence of the IL-1 family signature motif and the absence of a conventional ICE cut site. Phylogenetically, the grouper IL-1beta clustered closely with those of teleost belonging to Perciformes and apart from those of mammals. The grouper IL-1beta was constitutively expressed in almost all tissues examined, and was augmented in PBL after the addition of LPS or Poly I:C in vitro. The prokaryotically produced rIL-1beta significantly stimulated the proliferation of grouper head kidney cells, and activated gene expression of IL-1beta and COX-2. Moreover, the rIL-1beta-induced IL-1beta and COX-2 expression were reduced by p38 MAPK inhibitor (SB203580) and JNK inhibitor (SP600125), respectively. Taken together, the present study indicated that grouper IL-1beta may have an important role in grouper immune system and activate similar downstream cascades as its mammalian counterparts.
Lu
Dan-Qi
DQ
State Key Laboratory of Biocontrol, Institute of Aquatic Economic Animals and Guangdong Provincial Key Laboratory for Aquatic Economic Animals, College of Life Sciences, Sun Yat-Sen (Zhongshan) University, Guangzhou 510275, PR China.
Bei
Jin-Xin
JX
Feng
Li-Na
LN
Zhang
Yong
Y
Liu
Xiao-Chun
XC
Wang
Lei
L
Chen
Jie-Lin
JL
Lin
Hao-Ran
HR
D000595:Amino Acid Sequence; D000818:Animals; D000873:Anthracenes / Q000494:pharmacology; D001483:Base Sequence; D049109:Cell Proliferation; D002478:Cells, Cultured; D003001:Cloning, Molecular; D051546:Cyclooxygenase 2 / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D005786:Gene Expression Regulation; D007093:Imidazoles / Q000494:pharmacology; D053583:Interleukin-1beta / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D048670:MAP Kinase Kinase 4 / Q000037:antagonists & inhibitors / Q000378:metabolism; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010473:Perciformes / Q000235:genetics / Q000502:physiology*; D010802:Phylogeny; D011070:Poly I-C / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D015398:Signal Transduction; D048051:p38 Mitogen-Activated Protein Kinases
D000873:Anthracenes; D018076:DNA, Complementary; D007093:Imidazoles; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D011725:Pyridines; D011994:Recombinant Proteins; C432165:pyrazolanthrone; D051546:Cyclooxygenase 2; D048051:p38 Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4; D011070:Poly I-C; C093642:SB 203580