Abstract
The present study was performed to clone and characterize the structures and functions of steroidogenic factor 1 (sf-1) and 17α-hydroxylase/lyase (cyp17α) promoters in yellow catfish Pelteobagrus fulvidraco, a widely distributed freshwater teleost. We successfully obtained 1981 and 2034 bp sequences of sf-1 and cyp17α promoters, and predicted the putative binding sites of several transcription factors, such as Peroxisome proliferator-activated receptor alpha (PPARα), Peroxisome proliferator-activated receptor gamma (PPARγ) and Signal transducer and activator of transcription 3 (STAT3), on sf-1 and cyp17α promoter regions, respectively. Overexpression of PPARγ significantly increased the activities of sf-1 and cyp17α promoters, but overexpression of PPARα significantly decreased the promoter activities of sf-1 and cyp17α. Overexpression of STAT3 reduced the activity of the sf-1 promoter but increased the activity of the cyp17α promoter. The analysis of site-mutation and electrophoretic mobility shift assay suggested that the sf-1 promoter possessed the STAT3 binding site, but did not the PPARα or PPARγ binding sites. In contrast, only the PPARγ site, not PPARα or STAT3 sites, was functional with the cyp17α promoter. Leptin significantly increased sf-1 promoter activity, but the mutation of STAT3 and PPARγ sites decreased leptin-induced activation of sf-1 promoter. Our findings offered the novel insights into the transcriptional regulation of sf-1 and cyp17α and suggested leptin regulated sf-1 promoter activity through STAT3 site in yellow catfish.
Highlights
Steroid hormones are involved in the regulation of many physiological processes, such as embryonic development, sex differentiation, metabolism and reproduction in vertebrates [1], and their biosynthesis was regulated by key transcription factors and enzymes, such as steroidogenic factor 1 and 17α-hydroxylase [2]
On the cyp17α promoter, we predicted one TATA box located from −150 bp to −136 bp, one GC box located at −55 bp to −42 bp and one specificity protein 1 (SP1) binding site located at −53 bp to
Studies have indicated that the transcription of steroidogenic enzymes and steroid hormone receptors were mediated by transcription factor PPARs and Signal transducer and activator of transcription 3 (STAT3) [12,26], but the direct molecular evidences were absent
Summary
Steroid hormones are involved in the regulation of many physiological processes, such as embryonic development, sex differentiation, metabolism and reproduction in vertebrates [1], and their biosynthesis was regulated by key transcription factors and enzymes, such as steroidogenic factor 1 (sf-1) and 17α-hydroxylase (cyp17α) [2]. Several transcriptional factors, such as nuclear transcription factor Y (NF-Y), specificity protein 1 (SP1) and cAMP-response element binding protein (CREB), positively regulate the activities of sf-1 promoter [7,8,9]. Peroxisome proliferator-activated receptor alpha and gamma (PPARα and PPARγ) are the two important transcription factors that modulate the expression of many target genes involved in numerous physiological processes, including steroidogenesis [10,11,12]
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