title: Isolation and characterization of cyp19a1a and cyp19a1b promoters in the protogynous hermaphrodite orange-spotted grouper (Epinephelus coioides).
abstract: Aromatase (CYP19A1) catalyzes the conversion of androgens to estrogens. In teleosts, duplicated copies of cyp19a1 genes, namely cyp19a1a and cyp19a1b, were identified, however, the transcriptional regulation of these two genes remains poorly understood. In the present study, the 5'-flanking regions of the orange-spotted grouper cyp19a1a (gcyp19a1a) and cyp19a1b (gcyp19a1b) genes were isolated and characterized. The proximal promoter regions of both genes were relatively conserved when compared to those of the other teleosts. Notably, a conserved FOXO transcriptional factor binding site was firstly reported in the proximal promoter of gcyp19a1a, and deletion of the region (-112 to -60) containing this site significantly decreased the promoter activities. The deletion of the region (-246 to -112) containing the two conserved FTZ-F1 sites also dramatically decreased the transcriptional activities of gcyp19a1a promoter, and both two FTZ-F1 sites were shown to be stimulatory cis-acting elements. A FTZ-F1 homologue isolated from ricefield eel (eFTZ-F1) up-regulated gcyp19a1a promoter activities possibly via the FTZ-F1 sites, however, a previously identified orange-spotted grouper FTZ-F1 homologue (gFTZ-F1) did not activate the transcription of gcyp19a1a promoter unexpectedly. As to gcyp19a1b promoter, all the deletion constructs did not show good promoter activities in either TM4 or U251-MG cells. Estradiol (100nM) up-regulated gcyp19a1b promoter activities by about 13- and 36-fold in TM4 and U251-MG cells, respectively, via the conserved ERE motif, but did not stimulate gcyp19a1a promoter activities. These results are helpful to further elucidate the regulatory mechanisms of cyp19a1a and cyp19a1b expression in the orange-spotted grouper as well as other teleosts.
authors.affiliation: School of Life Sciences, Sun Yat-Sen University, Guanghzhou 510275, PR China. lsszwm@mail.sysu.edu.cn
authors.forename: Weimin
authors.initials: W
authors.lastname: Zhang
authors.forename: Huijie
authors.initials: H
authors.lastname: Lu
authors.forename: Haiyan
authors.initials: H
authors.lastname: Jiang
authors.forename: Mu
authors.initials: M
authors.lastname: Li
authors.forename: Shen
authors.initials: S
authors.lastname: Zhang
authors.forename: Qiongyou
authors.initials: Q
authors.lastname: Liu
authors.forename: Lihong
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001141:Aromatase / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001492:Bass / Q000378:metabolism*; D001665:Binding Sites; D005260:Female; D057850:Hermaphroditic Organisms / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D012741:Sexual Maturation / Q000502:physiology; D054339:Steroidogenic Factor 1 / Q000378:metabolism
chemical_list: D054339:Steroidogenic Factor 1; D001141:Aromatase