Functional Analysis of Promoters of Genes in Lipid Metabolism and Their Transcriptional Response to STAT3 under Leptin Signals

Wu, Kun-Lung; Tan, Xiao-Ying; Xu, Yi-Huan; Chen, Guang-Hui; Zhuo, Mei-Qin

doi:10.3390/genes9070334

Cited by 16 publications

(9 citation statements)

References 42 publications

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“…However, as far as we know, fish leptin is hardly available commercially at present. Therefore, recombinant human leptin has widely been used to investigate the function of fish leptin [ 22 , 23 , 37 , 38 ]. Thus, in the present study, recombinant human leptin was used to study promoters function of sf-1 and cyp17α in yellow catfish.…”

Section: Discussionmentioning

confidence: 99%

“…For example, several studies reported that leptin attenuated gonadotropin or growth factor-stimulated steroidogenesis in isolated theca and granulosa cells [ 17 , 18 ], but other studies pointed out that leptin induced the synthesis of estrogen in ovary and ovarian follicles [ 19 , 20 ]. Signal transducer and activator of transcription 3 (STAT3) is identified as the most important factor to transduce the leptin signal [ 21 , 22 , 23 ]. In mammals, studies have suggested that leptin modulated steroidogenic by STAT3 [ 13 , 19 ].…”

Section: Introductionmentioning

confidence: 99%

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

Chen

Zhuo

et al. 2020

IJMS

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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.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

Chen

Zhuo

et al. 2020

IJMS

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“…The hiTAIL-PCR (high-efficiency thermal asymmetric interlaced PCR) method [ 28 ] was used to clone the promoter sequences, and the protocols followed our previous studies [ 29 , 30 ]. The specific primers were listed in the Supplementary Table S1 .…”

Section: Methodsmentioning

confidence: 99%

“…Plasmid transfections and activity assays of luciferase followed our recent publications [ 29 , 30 ]. Briefly, HepG2 cells were cultured in DMEM medium (10% FBS) in an incubator with 5% CO 2 at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

Structural and Functional Analysis of SHP Promoter and Its Transcriptional Response to FXR in Zn-Induced Changes to Lipid Metabolism

Gao

Fan

et al. 2022

IJMS

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Zinc alleviates hepatic lipid deposition, but the transcriptional regulatory mechanisms are still unclear. In this study, we characterized the promoter of an SHP (short heterodimer partner) in a teleost Pelteobagrus fulvidraco. The binding sites of an FXR (farnesoid X receptor) were predicted by the SHP promoter, indicating that the FXR mediated its transcriptional activity. The site mutagenesis and the EMSA (electrophoretic mobility shift assay) found that the −375/−384 bp FXR site on the SHP promoter was the functional binding locus responsible for the Zn-induced transcriptional activation. A further study of yellow catfish hepatocytes suggested that the activation of the FXR/SHP is responsible for the effect of Zn on the decreasing lipid content. Thus, this study provides direct evidence of the interaction between the FXR and SHP promoter in fish, and accordingly elucidates the potential transcriptional mechanism by which Zn reduces hepatic lipid accumulation.

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“…For promoter luciferase assays, HepG2 cells were counted and seeded at a density of 1 × 10 5 in twenty-four well plates, then cultured and transfected as mentioned in our recent studies (19,23) . Briefly, to study the nSREBP-1-induced changes in promoter activities, we co-transfected 300 ng of nSREBP-1 plasmid or the same amount of pcDNA3.1(þ) plasmid (300 ng, control) with 500 ng of each of these luciferase reporter plasmids of srebp-1, accα, scd1 and fas promoters into HepG2 cells using Lipofectamine 2000 (Invitrogen) at 80-90 % confluence, respectively.…”

Section: Luciferase Assay Of Srebp-1 Accα Scd1 and Fas Promotersmentioning

confidence: 99%

Novel insights for SREBP-1 as a key transcription factor in regulating lipogenesis in a freshwater teleost, grass carp Ctenopharyngodon idella

Tan

et al. 2019

Br J Nutr

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Disturbances in lipid metabolism are at the core of several health issues facing modern society, including fatty liver and obesity. The sterol regulatory element-binding protein 1 (SREBP-1) is one important transcription factor regulating lipid metabolism, but the relevant mechanism still remains unknown. The present study determined the transcriptional regulation of SREBP-1 and its target genes (including acetyl-CoA carboxylase α (accα), fatty acid synthase (fas) and stearoyl-CoA desaturase 1 (scd1)) in a freshwater teleost, grass carp Ctenopharyngodon idella. We cloned and characterised the 1988 bp, 2043 bp, 1632 bp and 1889 bp sequences of srebp-1, accα, scd1 and fas promoters, respectively. A cluster of putative binding sites of transcription factors, such as specific protein, yin yang 1, nuclear factor Y, sterol response elements (SRE) and enhancer box (E-box) element, were predicted on their promoter regions. Overexpression of nSREBP-1 reduced srebp-1 promoter activity, increased scd1 and fas promoter activity but did not influence accα promoter activity. The site-mutation and electrophoretic mobility shift assay analysis indicated that srebp-1, fas and scd1 promoters, but not accα promoter, possessed SRE. In Ctenopharyngodon idella kidney (CIK) cells of grass carp, nSREBP-1 overexpression significantly reduced srebp-1 mRNA expression and up-regulated miR-29 mRNA expression. The 3′UTR of srebp-1 possessed the potential miR-29 binding site and miR-29 up-regulated the luciferase activity of srebp-1 3′UTR and srebp-1 mRNA expression, implying a self-activating loop of SREBP-1 and miR-29 in grass carp. Based on the above-mentioned results, we found two novel transcriptional mechanisms for SREBP-1 in grass carp: (1) the auto-regulation sited on the SREBP-1 promoter regions was suppressive and (2) there was a self-activating loop of SREBP-1 and miR-29.

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Functional Analysis of Promoters of Genes in Lipid Metabolism and Their Transcriptional Response to STAT3 under Leptin Signals

Cited by 16 publications

References 42 publications

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

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

Structural and Functional Analysis of SHP Promoter and Its Transcriptional Response to FXR in Zn-Induced Changes to Lipid Metabolism

Novel insights for SREBP-1 as a key transcription factor in regulating lipogenesis in a freshwater teleost, grass carp Ctenopharyngodon idella

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