title: Molecular characterizations and functional assessments of GATA-3 and its splice variant in Atlantic cod (Gadus morhua L.).
abstract: GATA-3 is a master transcription factor of the Th2 cells. We have identified GATA-3 cDNA and its splice variant in Atlantic cod. Cod GATA-3 (GmGATA-3) has a 1320 b p open reading frame encoding a polypeptide of 440 amino acids with two zinc finger domains that are well conserved within teleosts and higher vertebrates. The GATA-3 cDNA splice variant without zinc finger domains was shown to contain an 828 b p open reading frame encoding a polypeptide of 276 amino acids. Both GATA-3 proteins fused with RFP-tag were identified in or close to the nuclei 48 h after the plasmids were transfected in CHSE-214 cells. The full length GATA-3 with two zinc finger domains has a transcriptional function confirmed by transfection with GATA-3 reporter vector along with expression constructs of GATA-3 plasmids in CHSE-214 cells, whereas the GATA-3 splice variant without zinc finger domain did not enhance the activity of the GATA-3 reporter vector, and no interference was found between these two GATA-3 variants. RT-PCR analysis revealed that the two Atlantic cod GATA-3 variants were strongly expressed in the gills and infection with live Vibrio anguillarum induced the spleen expression of both GmGATA-3L and GmGATA-3S. Unexpectedly, PMA increased the expression of the GATA-3 splice variant in vivo and especially in vitro, with an increase of more than 100,000-fold in head kidney leukocytes at 24 and 48 h. On the other hand, there were no significant increases at the transcript level of full length GATA-3 between Poly I:C and β-glucan treatment groups compared to controls.
authors: Norwegian College of Fishery Science, University of Tromsø, N-9037 Tromsø, Norway.
authors: Heng
authors: H
authors: Chi
authors: Zuobing
authors: Z
authors: Zhang
authors: Makoto
authors: M
authors: Inami
authors: Jarl
authors: J
authors: Bøgwald
authors: Wenbin
authors: W
authors: Zhan
authors: Roy A
authors: RA
authors: Dalmo
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D029941:Fish Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D050990:GATA3 Transcription Factor / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D048389:Gadus morhua / Q000378:metabolism*; D060226:Head Kidney / Q000378:metabolism; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D016415:Sequence Alignment; D013154:Spleen / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D029941:Fish Proteins; D050990:GATA3 Transcription Factor; D020033:Protein Isoforms; D012333:RNA, Messenger