TAILIEUCHUNG - Báo cáo khoa học: Genomic structure, expression and characterization of a STAT5 homologue from pufferfish (Tetraodon fluviatilis)

TheSTAT5(signal transducer andactivator of transcription 5) genewas isolated and characterized froma round-spotted pufferfish genomic library. This gene is composed of 19 exons spanning 11 kb. The full-length cDNA ofTetraodon fluviatilis STAT5(TfSTAT5) contains 2461 bp and encodes a protein of 785 amino acid residues. From the amino acid sequence comparison,TfSTAT5 is most similar to mouse STAT5a and STAT5bwith an overall identity of 76% and 78%, respectively, and has | Eur. J. Biochem. 270 239-252 2003 FEBS 2003 doi Genomic structure expression and characterization of a STAT5 homologue from pufferfish Tetraodon fluviatilis k k B k B k Bk 1 B p B k Bk I OB OB B k 1 k B k 1 B B k Bk k B B 3 I B kk B k B k I I k BB B k BB I BB B B 1 kk I B II B B B 4 k B B B p B B B B k Bk k BB B k 1 Shu-Chiun Sung Ting-Jia Fan Chih-Ming Chou Jiann-Horng Leu Ya-Li Hsu Shui-Tsung Chen Yueh-Chun Hsieh1 and Chang-Jen Huang1 1Graduate Institute of Life Science National Defense Medical Center Taipei Taiwan 2Institute of Biological Chemistry and 4Zoology Academia Sinica Taipei Taiwan 3Department of Biochemistry Taipei Medical University Taipei Taiwan The STAT5 signal transducer and activator of transcription 5 gene was isolated and characterized from a round-spotted pufferfish genomic library. This gene is composed of 19 exons spanning 11 kb. The full-length cDNA of Tetraodon fluviatilis STAT5 fĩfSTAT5 contains 2461 bp and encodes a protein of 785 amino acid residues. From the amino acid sequence comparison TSTAT5 is most similar to mouse STAT5a and STAT5b with an overall identity of 76 and 78 respectively and has 35 identity with other mammalian STATs. The exon intron junctions of the TfSTAT5 gene were almost identical to those of mouse STAT5a and STAT5b genes indicating that these genes are highly conserved at the levels of amino acid sequence and genomic structure. To understand better the biochemical properties of TfSTAT5 a chimeric STAT5 was generated by fusion of the kinase-catalytic domain of carp Janus kinase 1 JAK1 to the C-terminal end of TfSTAT5. The fusion protein was expressed and tyrosine-phosphorylated by its kinase domain. The fusion protein exhibits specific DNA-binding and transactivation potential toward an artificial fish promoter as well as authentic mammalian promoters such as the b-casein promoter and cytokine inducible SH2 containing protein CIS promoter when expressed in both fish and mammalian .

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