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      CCAAT-enhancer binding protein-α (C/EBPα) and hepatocyte nuclear factor 4α (HNF4α) regulate expression of the human fructose-1,6-bisphosphatase 1 (FBP1) gene in human hepatocellular carcinoma HepG2 cells

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          Abstract

          Fructose-1,6-bisphosphatase (FBP1) plays an essential role in gluconeogenesis. Here we report that the human FBP1 gene is regulated by two liver-enriched transcription factors, CCAAT-enhancer binding protein-α (C/EBPα) and hepatocyte nuclear factor 4α (HNF4α) in human hepatoma HepG2 cells. C/EBPα regulates transcription of FBP1 gene via binding to the two overlapping C/EBPα sites located at nucleotide -228/-208 while HNF4α regulates FBP1 gene through binding to the classical H4-SBM site and direct repeat 3 (DR3) located at nucleotides -566/-554 and -212/-198, respectively. Mutations of these transcription factor binding sites result in marked decrease of C/EBPα- or HNF4α-mediated transcription activation of FBP1 promoter-luciferase reporter expression. Electrophoretic mobility shift assays of -228/-208 C/EBPα or -566/-554 and -212/-198 HNF4α sites with nuclear extract of HepG2 cells overexpressing C/EBPα or HNF4α confirms binding of these two transcription factors to these sites. Finally, we showed that siRNA-mediated suppression of C/EBPα or HNF4α expression in HepG2 cells lowers expression of FBP1 in parallel with down-regulation of expression of other gluconeogenic enzymes. Our results suggest that an overall gluconeogenic program is regulated by these two transcription factors, enabling transcription to occur in a liver-specific manner.

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          Most cited references55

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          PROMO: detection of known transcription regulatory elements using species-tailored searches.

          We have developed a set of tools to construct positional weight matrices from known transcription factor binding sites in a species or taxon-specific manner, and to search for matches in DNA sequences.
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            The nuclear receptor superfamily: the second decade.

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              Loss of FBP1 by Snail-mediated repression provides metabolic advantages in basal-like breast cancer.

              The epithelial-mesenchymal transition (EMT) enhances cancer invasiveness and confers tumor cells with cancer stem cell (CSC)-like characteristics. We show that the Snail-G9a-Dnmt1 complex, which is critical for E-cadherin promoter silencing, is also required for the promoter methylation of fructose-1,6-biphosphatase (FBP1) in basal-like breast cancer (BLBC). Loss of FBP1 induces glycolysis and results in increased glucose uptake, macromolecule biosynthesis, formation of tetrameric PKM2, and maintenance of ATP production under hypoxia. Loss of FBP1 also inhibits oxygen consumption and reactive oxygen species production by suppressing mitochondrial complex I activity; this metabolic reprogramming results in an increased CSC-like property and tumorigenicity by enhancing the interaction of β-catenin with T-cell factor. Our study indicates that the loss of FBP1 is a critical oncogenic event in EMT and BLBC. Copyright © 2013 Elsevier Inc. All rights reserved.

                Author and article information

                Contributors
                Role: ConceptualizationRole: InvestigationRole: Writing – original draft
                Role: Supervision
                Role: Data curationRole: Investigation
                Role: Supervision
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: Supervision
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                22 March 2018
                2018
                : 13
                : 3
                : e0194252
                Affiliations
                [1 ] Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, Thailand
                [2 ] Division of Interdisciplinary, Mahidol University, Kanjanaburi, Thailand
                [3 ] Childrens Diabetes Center, University of Wisconsin School of Medicine and Public Health, Madison, WI, United States of America
                Rosalind Franklin University of Medicine and Science, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0002-5878-6291
                Article
                PONE-D-18-01514
                10.1371/journal.pone.0194252
                5863999
                29566023
                07d74ef9-4233-4e12-868d-f78fea6a8253

                This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

                History
                : 16 January 2018
                : 27 February 2018
                Page count
                Figures: 6, Tables: 1, Pages: 20
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100004396, Thailand Research Fund;
                Award ID: IRN59W0003
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100004396, Thailand Research Fund;
                Award ID: RGJ-PHD scholarship (4CMU56010)
                Award Recipient :
                This work was supported by an international research network (grant IRN 59W0003), the Thailand Research Fund. S.W. was supported by an RGJ-PHD scholarship (4CMU56010), the Thailand Research Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Cell Biology
                Cell Physiology
                Cell Binding
                Biology and life sciences
                Biochemistry
                Proteins
                DNA-binding proteins
                Transcription Factors
                Biology and Life Sciences
                Genetics
                Gene Expression
                Gene Regulation
                Transcription Factors
                Biology and Life Sciences
                Biochemistry
                Proteins
                Regulatory Proteins
                Transcription Factors
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
                Gene Mapping
                Restriction Fragment Mapping
                Electrophoretic Mobility Shift Assay
                Research and Analysis Methods
                Molecular Biology Techniques
                Gene Mapping
                Restriction Fragment Mapping
                Electrophoretic Mobility Shift Assay
                Biology and Life Sciences
                Genetics
                Gene Expression
                Gene Regulation
                Transcriptional Control
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
                Sequencing Techniques
                Nucleotide Sequencing
                Research and Analysis Methods
                Molecular Biology Techniques
                Sequencing Techniques
                Nucleotide Sequencing
                Biology and Life Sciences
                Biochemistry
                Enzymology
                Enzymes
                Oxidoreductases
                Luciferase
                Biology and Life Sciences
                Biochemistry
                Proteins
                Enzymes
                Oxidoreductases
                Luciferase
                Biology and life sciences
                Genetics
                Gene expression
                DNA transcription
                Biology and Life Sciences
                Cell Biology
                Cellular Types
                Animal Cells
                Hepatocytes
                Biology and Life Sciences
                Anatomy
                Liver
                Hepatocytes
                Medicine and Health Sciences
                Anatomy
                Liver
                Hepatocytes
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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