Functional analysis of MTF-1 and MT promoters and their transcriptional response to zinc (Zn) and copper (Cu) in yellow catfish Pelteobagrus fulvidraco

Chen, Guanghui; Lv, Wu-Hong; Xu, Yi-Huan; Wei, Xiaolei

doi:10.1016/j.chemosphere.2019.125792

Cited by 31 publications

(32 citation statements)

References 37 publications

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“…Further EMSA indicated that the −1543 bp/−1557 bp MTF-1 binding site could be bound by MTF-1, and that Zn promoted the binding of MTF-1 to its binding site on the ZIP8 promoter. The indispensable role of Zn-MTF-1 axis in regulating Zn homeostasis has been mentioned in several reports [25,26], and that fish and mammalian cells could respond to Zn 2+ by increasing the MTF-1 binding [8,27]. Andrews [17] pointed out that the MTF-1 binding to the MRE was dependent on Zn and was easily disrupted when Zn was depleted.…”

Section: Discussionmentioning

confidence: 99%

“…Similarly, one MTF-1 binding site were found in this region. MTF-1 was reported to positively regulate ZIP family [7,8].…”

Section: Discussionmentioning

confidence: 99%

“…Further site mutagenesis and EMSA identified a functional binding site of MTF-1 in these regions. MTF-1 functions as a cellular Zn sensor and binds specifically to MREs (metal response elements) to activate the expression of genes encoding Zn transporters in response to Zn ions [8,17,18]. Ling et al [19] reported that 0.18 mg/L of waterborne Zn addition could significantly improve the mRNA expression level of MTF-1 of juvenile goby Synechogobius hasta.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, as the first step for demonstrating the regulatory mechanism of Zn transporters, it is very important to investigate the structure and function of the promoters. In addition, metal-regulatory transcription factor-1 (MTF1) and ras-responsive element-binding protein 1 (RREB1) are two important Zn-finger (Cys2His2) transcription factors and function as sensors of intracellular Zn by binding to their corresponding responsive elements, and thereby regulates cellular adaptation to Zn [6][7][8]. At present, the responsive elements of MTF-1 and RREB1 have been identified in the promoter regions of several Zn transporters [6,8], but not in ZIP3 and ZIP8.…”

Section: Introductionmentioning

confidence: 99%

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Functional Analysis of Two Zinc (Zn) Transporters (ZIP3 and ZIP8) Promoters and Their Distinct Response to MTF1 and RREB1 in the Regulation of Zn Metabolism

Chen

et al. 2020

IJMS

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ZIP (zinc-regulated transporters, iron-regulated transporter-like protein) family plays an important role in organism Zn balance. This research identified the promoter regions of ZIP3 and ZIP8, two members of ZIP family, from a freshwater teleost yellow catfish Pelteobagrus fulvidraco, characterized the binding sequences of the metal-responsive transcription factor-1 (MTF-1) and Ras responsive element binding protein 1 (RREB1) on their promoter regions. The present study cloned and obtained the 2027 bp of ZIP3 promoter and 1664 bp of ZIP8 promoter, and predicted several key elements on their promoters, such as the binding sites of CREB (cAMP-response element binding protein), KLF4 (Kruppel like factor 4), MTF-1 and RREB1. The sequence deletion from −361 bp to −895 bp down-regulated the luciferase activity of ZIP3 promoter, and the deletion from −897 bp to −1664 bp down-regulated the luciferase activity of ZIP8 promoter. Within different deletion plasmids, the relative luciferase activities of ZIP3 and ZIP8 promoters changes to Zn incubation in a Zn concentration-dependent manner. The site mutagenesis and EMSA (electrophoretic mobility shift assay) found that the −1327 bp/−1343 bp MTF-1 binding site and the −248 bp/−267 bp RREB1 binding site on the ZIP3 promoter, and the −1543 bp/−1557 bp MTF-1 binding site on the ZIP8 promoter are functional sites. Low Zn increased the binding capability between MTF-1 and its responsive site on the ZIP3 promoter, and high Zn increased the transcriptional activation ZIP3 by RREB1; Zn also promoted the binding ability between MTF-1 and its responsive element on the ZIP8 promoter. This study provides the first direct evidence for the response elements of MTF-1 and RREB1 on ZIP3 and MTF-1 on ZIP8 to Zn, which are very important for the evaluation of Zn nutrition and toxicity in vertebrates.

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Section: Discussionmentioning

confidence: 99%

“…Similarly, one MTF-1 binding site were found in this region. MTF-1 was reported to positively regulate ZIP family [7,8].…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Functional Analysis of Two Zinc (Zn) Transporters (ZIP3 and ZIP8) Promoters and Their Distinct Response to MTF1 and RREB1 in the Regulation of Zn Metabolism

Chen

et al. 2020

IJMS

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“…The gene expression was determined using the RT-qPCR method as reported by Chen et al [ 22 ], and microRNAs expression was assayed by the method of stem–loop RT–PCR [ 16 ]. The specific stem-loop primers and PCR primers for each gene are shown in Table S1 .…”

Section: Methodsmentioning

confidence: 99%

miR-101b Regulates Lipid Deposition and Metabolism of Primary Hepatocytes in Teleost Yellow Catfish Pelteobagrus fulvidraco

Chen

Zhao

Wei

et al. 2020

Genes

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Excessive fat deposition in the hepatocytes, associated with excess dietary fat intake, was related to the occurrence of fatty livers in fish. miR-101b plays the important roles in controlling lipid metabolism, but the underlying mechanism at the post-transcriptional level remains unclear. The purpose of this study is to explore the roles and mechanism of miR-101b-mediating lipid deposition and metabolism in yellow catfish Pelteobagrus fulvidraco. We found that miR-101b directly targeted fatty acid translocase (cd36), caspase9 (casp9) and autophagy-related gene 4A (atg4a). Furthermore, using palmitic acid (PA) or oleic acid (OA) to incubate the primary hepatocytes of yellow catfish, we demonstrated that miR-101b inversely regulated cd36, casp9, and atg4a expression at the transcriptional level; the inhibition of miR-101b aggravated fatty acids (FAs, PA or OA)-induced lipid accumulation, indicating that miR-101b mediated FAs-induced variations of lipid metabolism in yellow catfish. Taken together, our study gave novel insight into the regulatory mechanism of lipid deposition and metabolism and might provide potential targets for the prevention and treatment of fatty livers in fish.

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

Xu,

He,

Liang

et al. 2023

Journal Cellular Physiology

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The melanocortin 4 receptor (MC4R) is a G protein‐coupled transporter that mediates the regulation of thyroid hormones and leptin on energy balance and food intake. However, the mechanisms of transcriptional regulation of Mc4r by thyroid hormone and leptin in fish have been rarely reported. The messenger RNA expression of Mc4r gene was significantly higher in brain than those in other tissues of mandarin fish. We analyzed the structure and function of a 2029 bp sequence of Mc4r promoter. Meanwhile, overexpression of NKX2.1 and incubation with leptin significantly increased Mc4r promoter activity, but triiodothyronine showed the opposite effect. In addition, mutations in the NKX2.1 binding site abolished not only the activation of Mc4r promoter activity by leptin but also the inhibitory effect of thyroid hormones on Mc4r promoter activity. In summary, these results suggested that thyroid hormones and leptin might regulate the transcriptional expression of Mc4r through NKX2.1.

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Functional analysis of MTF-1 and MT promoters and their transcriptional response to zinc (Zn) and copper (Cu) in yellow catfish Pelteobagrus fulvidraco

Cited by 31 publications

References 37 publications

Functional Analysis of Two Zinc (Zn) Transporters (ZIP3 and ZIP8) Promoters and Their Distinct Response to MTF1 and RREB1 in the Regulation of Zn Metabolism

Functional Analysis of Two Zinc (Zn) Transporters (ZIP3 and ZIP8) Promoters and Their Distinct Response to MTF1 and RREB1 in the Regulation of Zn Metabolism

miR-101b Regulates Lipid Deposition and Metabolism of Primary Hepatocytes in Teleost Yellow Catfish Pelteobagrus fulvidraco

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

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