ZFIN ID: ZDB-PUB-090424-20
Organization and promoter analysis of the zebrafish (Danio rerio) chemokine gene (CXC-64) promoter
Chen, L.C., Wu, J.L., Shiau, C.Y., and Chen, J.Y.
Date: 2010
Source: Fish physiology and biochemistry   36(3): 511-521 (Journal)
Registered Authors: Chen, Jyh-Yih, Wu, Jen-Leih
Keywords: Zebrafish, Chemokine, Gene expression, Functional analysis
MeSH Terms:
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Line, Tumor
  • Chemokines, CXC/genetics*
  • Cloning, Molecular
  • DNA Primers/genetics
  • Gene Components
  • Gene Expression Regulation/genetics*
  • Humans
  • Lipopolysaccharides/metabolism
  • Mice
  • Molecular Sequence Data
  • Promoter Regions, Genetic/genetics*
  • Receptors, Cell Surface/metabolism
  • Sequence Analysis, DNA
  • Tumor Necrosis Factor-alpha/metabolism
  • Zebrafish/genetics*
PubMed: 19381848 Full text @ Fish Physiol. Biochem.
ABSTRACT
Zebrafish CXC-64, a chemokine representing a superfamily of chemotactic cytokines present in fish, is involved in recruitment, activation, and response to inflammatory stimulation. We cloned and sequenced the genomic DNA of the zebrafish CXC-64 chemokine; it was most similar to CXCL11 from humans and CXCL10 from a catfish. The zebrafish CXC-64 gene is approximately 4.0 kb long and has a four-exon, three-intron structure common to the human CXCL11 gene. However, the promoter region includes a typical TATA box and multi-transcription factor-binding sequences. To understand the roles of lipopolysaccharide (LPS), poly I:poly C, and tumor necrosis factor (TNF)-alpha in regulating zebrafish CXC-64 expression, serial deletions were made in the promoter region of this clone. Different fragments of the zebrafish CXC-64 5'-flanking region were transfected into RAW264.7 (mouse macrophage; Abelson murine leukemia virus transformed) and zfl (zebrafish liver) cells and then treated with 0, 10, 50, 100, and 200 ng/ml LPS, poly I:poly C, or TNF-alpha. The results showed that the promoter activity presented dose-dependent effects in LPS-treated RAW264.7 cells, TNF-alpha-treated RAW264.7 cells, and LPS-treated zfl cells. These results reveal that the zebrafish CXC-64 chemokine gene promoter region can be induced by LPS in both human and fish cell lines, which suggests that it plays an important role in regulating LPS.
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