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

2021 ◽  
Vol 22 (9) ◽  
pp. 4505
Author(s):  
Fang Chen ◽  
Chong-Chao Zhong ◽  
Chang-Chun Song ◽  
Shu-Wei Chen ◽  
Yang He ◽  
...  
Keyword(s):  
Promoter Activity ◽  
Tissue Expression ◽  
Gonadal Development ◽  
Pelteobagrus Fulvidraco ◽  
The Novel ◽  
Yellow Catfish ◽  
Site Mutation ◽  
Promoter Regions ◽  
Promoter Sequences ◽  
Different Levels

The steroid hormones are required for gonadal development in fish. The present study was undertaken to characterize the cDNA and promoter sequences of TSPO and SMAD4 genes in yellow catfish Pelteobagrus fulvidraco, explored the mRNA tissue expression and deciphered their promoter regions. Yellow catfish TSPO and SMAD4 shared the similar domains to the corresponding genes from other vertebrates. The TSPO and SMAD4 mRNAs were widely expressed in the detected tissues, but at different levels. Several transcription factors were predicted, such as Sp, GATA, AP1, SOX1, SRY, STAT, HNF4α, PPARγ, Pu.1 and FOXL2. PPARγ overexpression increased but STAT3 overexpression reduced TSPO promoter activity, and FOXL2 overexpression inhibited the promoter activity of TSPO and SMAD4. The site mutation and EMSA analysis indicated that TSPO promoter possessed STAT3 and FOXL2 sites. Overall, our provided the novel understanding into the transcriptionally regulatory mechanisms of TSPO and SMAD4 in fish.

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

2020 ◽  
Vol 22 (1) ◽  
pp. 195
Author(s):  
Wu-Hong Lv ◽  
Guang-Hui Chen ◽  
Mei-Qin Zhuo ◽  
Yi-Huan Xu ◽  
Yi-Chuang Xu ◽  
...  
Keyword(s):  
Binding Sites ◽  
Promoter Activity ◽  
Pelteobagrus Fulvidraco ◽  
Peroxisome Proliferator ◽  
Yellow Catfish ◽  
Site Mutation ◽  
Promoter Regions ◽  
Mobility Shift ◽  
Peroxisome Proliferator Activated Receptor ◽  
Steroidogenic Factor 1

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.

scholarly journals Novel Insights for PI3KC3 in Mediating Lipid Accumulation in Yellow Catfish Pelteobagrus Fulvidraco

2021 ◽  
Author(s):  
Mei-Qin Zhuo ◽  
Jun Chen ◽  
Mei-Li Wu ◽  
Wen-biao Wang
Keyword(s):  
Transcriptional Regulation ◽  
Protein Level ◽  
Lipid Accumulation ◽  
Mrna Level ◽  
Pelteobagrus Fulvidraco ◽  
Yellow Catfish ◽  
Site Mutation ◽  
Mobility Shift ◽  
Mobility Shift Assay ◽  
Mutation Assay

Abstract In this study, the transcriptional regulation of PI3KC3 by three transcript factors (PPARγ, PPARα and STAT3) and the potential role of PI3KC3 in mediating lipid accumulation were determined in yellow catfish Pelteobagrus fulvidraco. The 5’-deletion assay, overexpression assay, site-mutation assay and electrophoretic mobility shift assay suggested that PPARα, PPARγ and STAT3 negatively regulated the promoter activity of pi3kc3. Moreover, the transcriptional inactivation of pi3kc3 was directly mediated by PPARα and PPARγ under fatty acid (FA) treatment. Using primary hepatocytes from yellow catfish, FA incubation significantly increased triacylglyceride (TG), NEFA content, the mRNA level of pparα, pparγ, stat3 and dnmt3b, the protein level of PPARα, PPARγ and STAT3, and the methylation level of pi3kc3, but significantly reduced the mRNA and protein level of PI3KC3. Our findings offer new insights into the mechanisms for transcriptional regulation of PI3KC3 and for PI3KC3-mediated lipid accumulation in fish.

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