Expression and characterization of the JAK kinase and STAT protein from brine shrimp, Artemia franciscana

Chia Hsiung Cheng, Gen Der Chen, Maw Sheng Yeh, Cheng Ying Chu, Ya Li Hsu, Pung Pung Hwang, Fore Lien Huang, Chang Jen Huang

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

In this study, we isolated and characterized both JAK and STAT genes from Artemia, Artemia franciscana. Although AfJAK showed only 19% identity (33% similarity) to the Drosophila Hop protein, AfJAK contained the characteristic JAK homology domain (JH domain) from JH1 to JH7. On the other hand, AfSTAT showed higher identity (30%) to Drosophila STAT (STAT92E). The low identities of AfJAK and AfSTAT to Drosophila Hop and STAT92E suggest that JAK and STAT proteins are unique in each different species of invertebrate. RT-PCR analysis showed that both AfJAK and AfSTAT transcripts were ubiquitously expressed in the embryo, which is similar to the expression patterns of Drosophila Hop and STAT92E mRNAs during development. In addition, we generated a constitutively active form of AfSTAT by fusing the JH1 domain of AfJAK to the C-terminal end of AfSTAT. This fusion protein, AfSTAT-HA-JH1, autophosphorylated on its tyrosine residue and was able to bind to specific DNA motifs including the STAT-binding motifs in the Drosophila Raf promoter. Both AfJAK and AfSTAT proteins elicited the transactivation potential toward the fly Raf promoter in Sf9 cells. However, tyrosine phosphorylation of AfSTAT was not detected, which is consistent with the cellular localization analysis that most AfSTAT proteins were in the cytoplasm. Our results demonstrate that both JAK and STAT are present in the genome of Artemia, which can serve as the basis for further investigations to explore the role of the JAK/STAT signal pathway in the development and immune response of brine shrimp.

Original languageEnglish
Pages (from-to)774-782
Number of pages9
JournalFish and Shellfish Immunology
Volume28
Issue number5-6
DOIs
Publication statusPublished - May 2010

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Janus Kinases
Artemia franciscana
Artemia
brine
Drosophila
phosphotransferases (kinases)
hops
protein
Tyrosine
Proteins
proteins
Genes
tyrosine
Phosphorylation
promoter regions
transcriptional activation
cytoplasm
immune response
sequence homology
homology

ASJC Scopus subject areas

  • Aquatic Science
  • Environmental Chemistry

Cite this

Expression and characterization of the JAK kinase and STAT protein from brine shrimp, Artemia franciscana. / Cheng, Chia Hsiung; Chen, Gen Der; Yeh, Maw Sheng; Chu, Cheng Ying; Hsu, Ya Li; Hwang, Pung Pung; Huang, Fore Lien; Huang, Chang Jen.

In: Fish and Shellfish Immunology, Vol. 28, No. 5-6, 05.2010, p. 774-782.

Research output: Contribution to journalArticle

Cheng, Chia Hsiung ; Chen, Gen Der ; Yeh, Maw Sheng ; Chu, Cheng Ying ; Hsu, Ya Li ; Hwang, Pung Pung ; Huang, Fore Lien ; Huang, Chang Jen. / Expression and characterization of the JAK kinase and STAT protein from brine shrimp, Artemia franciscana. In: Fish and Shellfish Immunology. 2010 ; Vol. 28, No. 5-6. pp. 774-782.
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