Expression and characterization of a constitutively active STAT6 from Tetraodon

Shu Chiun Sung, Chia Hsiung Cheng, Chih Ming Chou, Cheng Ying Chu, Gen Der Chen, Pung Pung Hwang, Fore Lien Huang, Chang Jen Huang

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

In this paper, we report the cloning and characterization of the STAT6 gene from the pufferfish, Tetraodon nigroviridis. The TnSTAT6 gene is composed of 20 exons and 19 introns. The exon-intron organization of this gene is similar to that of HsSTAT6 except for the exons encoding the C-terminal transactivation domain. The full-length complementary (c)DNA of TnSTAT6 encodes a 794-amino acid protein that is 31% identical to human STAT6. We generated a constitutively active TnSTAT6-JH1 by fusing the kinase domain of carp JAK1 to the C-terminal end of TnSTAT6 and demonstrated that the fusion protein has specific DNA-binding ability and can activate a reporter construct carrying multiple copies of mammalian IL-4-response elements. Interestingly, TnSTAT6-JH1 associated with and phosphorylated TnSTAT6 on Tyr661. Mutation of this residue, Y661W, in TnSTAT6 abolished its association with TnSTAT6-JH1. This is consistent with the importance of the corresponding Tyr641 of HsSTAT6 in tyrosine phosphorylation and dimer formation. On the other hand, treatment of mammalian IL-4 did not induce tyrosine phosphorylation of wild-type TnSTAT6, suggesting that both the divergent N-terminal domain and coiled-coiled domain of TnSTAT6 may affect the interaction of TnSTAT6 with mammalian IL-4 receptor complexes.

Original languageEnglish
Pages (from-to)819-828
Number of pages10
JournalFish and Shellfish Immunology
Volume28
Issue number5-6
DOIs
Publication statusPublished - May 2010

Fingerprint

interleukin-4
exons
Exons
Phosphorylation
Genes
Interleukin-4
Introns
Tyrosine
tyrosine
gene
introns
phosphorylation
Interleukin-4 Receptors
DNA
Tetraodontidae
protein
genes
Cloning
response elements
binding capacity

Keywords

  • DNA-binding
  • Interleukin 4
  • Pufferfish
  • Reporter assay
  • STAT6

ASJC Scopus subject areas

  • Aquatic Science
  • Environmental Chemistry

Cite this

Expression and characterization of a constitutively active STAT6 from Tetraodon. / Sung, Shu Chiun; Cheng, Chia Hsiung; Chou, Chih Ming; Chu, Cheng Ying; Chen, Gen Der; Hwang, Pung Pung; Huang, Fore Lien; Huang, Chang Jen.

In: Fish and Shellfish Immunology, Vol. 28, No. 5-6, 05.2010, p. 819-828.

Research output: Contribution to journalArticle

Sung, Shu Chiun ; Cheng, Chia Hsiung ; Chou, Chih Ming ; Chu, Cheng Ying ; Chen, Gen Der ; Hwang, Pung Pung ; Huang, Fore Lien ; Huang, Chang Jen. / Expression and characterization of a constitutively active STAT6 from Tetraodon. In: Fish and Shellfish Immunology. 2010 ; Vol. 28, No. 5-6. pp. 819-828.
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