Functional Analysis of Steroidogenic Factor 1 (sf-1) and 17α-Hydroxylase/Lyase (cyp17α) Promoters in Yellow Catfish Pelteobagrus fulvidraco

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.

Regulatory effect of NK homeobox 1 (NKX2.1) on melanocortin 4 receptor (Mc4r) promoter in Mandarin fish

“…Thus, 100 nM T3 was adopted in the current study (Figure 6a). To study the responsiveness of the Mc4r promoter to thyroxine and leptin, HEK293T cells were incubated with 100 nM T3 or 200 ng/mL leptin (Lv et al, 2020) for 24 h.…”

“…The optimal concentration of T3 was screened by the effect of T3 on the activity of the Mc4r promoter. After transfection of Mc4r promoter 5′‐deletion fragment plasmids (pGl3‐41/+321, pGl3‐282/+321, pGl3‐908/+321, and pGl3‐1708/+321), pcDNA3.1(+)‐NKX2.1, and Mut‐ Mc4r ‐NKX2.1, the transfected HEK293T cells were incubated using 0 and 200 ng/mL leptin (Lv et al, 2020), and optimal concentrations of T3, respectively. Collect cell samples after 24 h, the cells were lysed and harvested for the luciferase activities according to the procedure above.…”

Molecular Characterization and Functional Analysis of Two Steroidogenic Genes TSPO and SMAD4 in Yellow Catfish

“…PPARγ, STAT3 and FOXL2 regulate the transcription of key enzymes in steroidogenesis [ 13 , 38 ]. The present study found that PPARγ significantly increased the transcriptional activity of TSPO promoter, while STAT3 and FOXL2 decreased TSPO transcription.…”

“…Yellow catfish Pelteobagrus fulvidraco , widely distributed freshwater fish, is an important economic fish in some Asian countries due to the high economic value and delicious meat quality. At present, studies were scarce on the regulation of steroidogenesis for the fish species [ 13 ]. To this end, we cloned and characterized the cDNA and promoter sequences of TSPO and SMAD4 genes, and determined their mRNA tissue expression and investigated their transcriptional regulation in yellow catfish.…”

Novel insights for PI3KC3 in mediating lipid accumulation in yellow catfish Pelteobagrus fulvidraco

“…The primers for the plasmids construction were presented in Table S2. In addition, The overexpression plasmids of PPARα, PPARγ and STAT3 were obtained from our previous studies (Lv et al 2021).…”