Genomic structure and functional analysis of promoter region of somatolactin gene of sea bream (Sparus aurata).
Somatolactin (SL) is a pituitary hormone belonging to the growth hormone-prolactin family and is produced in the intermediate lobe of teleosts. The SL gene was isolated from a sea bream genomic library and found to be composed of 5 exons distributed within a 9-kb length of DNA. Sequence analysis of the proximal promoter region showed the presence of a classical TATA box located 59 bp upstream from the initial start ATG codon, 5 consensus sequences corresponding to the Pit-1 binding element, and a putative CREB site. In CHO cells cotransfected with the DNA from 2 plasmids, one encoding sea bream Pit-1 under Rous sarcoma virus long terminal repeat regulation and one encoding the SL promoter driving the expression of luciferase, Pit-1 was found to enhance the expression of luciferase. Only one Pit-1 binding site was necessary for enhancement. Analysis by immunoblots of in vitro culture of pituitaries of Sparus aurata showed that several agents, including estradiol, verapamil, and phorbol myristate acetate, had different inhibitory effects on SL and growth hormone released to the culture medium.
Astola
Antonio
A
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Cádiz, 11510 Puerto Real, Cádiz, Spain.
Calduch-Giner
Josep A
JA
Ortiz
Manuela
M
Pérez-Sánchez
Jaume
J
Valdivia
Manuel M
MM
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D004958:Estradiol / Q000494:pharmacology; D029941:Fish Proteins; D040461:Gene Components; D015698:Genomic Library; D006023:Glycoproteins / Q000235:genetics*; D015151:Immunoblotting; D008156:Luciferases / Q000378:metabolism; D008969:Molecular Sequence Data; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism; D010907:Pituitary Hormones / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D021541:Sea Bream / Q000235:genetics*; D017422:Sequence Analysis, DNA; D050608:Sodium-Phosphate Cotransporter Proteins; D027981:Symporters / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection; D014700:Verapamil / Q000494:pharmacology
D029941:Fish Proteins; D006023:Glycoproteins; D010907:Pituitary Hormones; D050608:Sodium-Phosphate Cotransporter Proteins; D027981:Symporters; C064448:somatolactin protein, fish; D004958:Estradiol; D014700:Verapamil; D008156:Luciferases; D013755:Tetradecanoylphorbol Acetate