SREBP-1c and Sp1 Interact to Regulate Transcription of the Gene for Phosphoenolpyruvate Carboxykinase (GTP) in the Liver*

The sterol regulatory element-binding protein-1c (SREBP-1c), as well as SREBP-1a and SREBP-2, inhibit transcription of the gene encoding the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) (PEPCK-C). There are two SREBP regulatory elements (SREs) in the PEPCK-C gene promoter (-322 to -313 and -590 to -581). The SRE at -590 overlaps an Sp1 site on the opposite strand of the DNA. These SREs bound SREBP-1a and SREBP-1c with low affinity but the addition of purified upstream stimulatory activity enhanced the binding of SREBP-1 to both of these sites. Mutating these SREs increased both unstimulated (5-fold) and protein kinase A-stimulated transcription (8-27-fold) from the PEPCK-C gene promoter; this was lost when both SREs were mutated. The SRE at -590 differs by a single base pair from the SRE in the low density lipoprotein (LDL) receptor gene (T in the PEPCK-C gene promoter at -582, compared with an A in the SRE of the gene for the LDL receptor promoter). Introduction of the LDL receptor SRE into the PEPCK-C gene promoter increased SREBP-1c binding and caused a 10-fold enhancement of basal transcription from the promoter, rather than an inhibition as observed with the SRE in the PEPCK-C gene promoter. The T/A change does not alter the binding of Sp1 to its site on the opposite strand of the DNA. Sp1 bound to the promoter independently of SREBP-1c but competed with SREBP-1c for binding. Sp1 does not bind to the SRE at -322. Chromatin immunoprecipitation analysis, using rat hepatocytes, demonstrated that SREBP-1 and Sp1 were associated in vivo with putative regulatory regions corresponding to the SREs in the PEPCK-C gene promoter. We propose that insulin represses transcription of the gene for PEPCK-C by inducing SREBP-1c production in the liver, which interferes with the stimulatory effect of Sp1 at -590 of the PEPCK-C gene promoter.

[1]  F. Bone,et al.  Regulation of Glucose-6-phosphatase Gene Expression in Cultured Hepatocytes and H4IIE Cells by Short-chain Fatty Acids , 2003, Journal of Biological Chemistry.

[2]  Bruce M. Spiegelman,et al.  Insulin-regulated hepatic gluconeogenesis through FOXO1–PGC-1α interaction , 2003, Nature.

[3]  Hitoshi Shimano,et al.  Transcriptional activities of nuclear SREBP-1a, -1c, and -2 to different target promoters of lipogenic and cholesterogenic genes DOI 10.1194/jlr.M100417-JLR200 , 2002, Journal of Lipid Research.

[4]  R. Hammer,et al.  Diminished Hepatic Response to Fasting/Refeeding and Liver X Receptor Agonists in Mice with Selective Deficiency of Sterol Regulatory Element-binding Protein-1c* , 2002, The Journal of Biological Chemistry.

[5]  D. Azzout-Marniche,et al.  Adenovirus-mediated overexpression of sterol regulatory element binding protein-1c mimics insulin effects on hepatic gene expression and glucose homeostasis in diabetic mice. , 2001, Diabetes.

[6]  H. Shimano.  Sterol regulatory element-binding proteins (SREBPs): transcriptional regulators of lipid synthetic genes. , 2001, Progress in lipid research.

[7]  M. Foretz,et al.  Sterol Regulatory Element-binding Protein-1c Mimics the Negative Effect of Insulin on Phosphoenolpyruvate Carboxykinase (GTP) Gene Transcription* , 2001, The Journal of Biological Chemistry.

[8]  Jianqi Yang,et al.  Sodium Butyrate Induces Transcription from the Gαi2Gene Promoter through Multiple Sp1 Sites in the Promoter and by Activating the MEK-ERK Signal Transduction Pathway* , 2001, The Journal of Biological Chemistry.

[9]  R. Hanson,et al.  The Use of Transgenic Mice to Analyze the Role of Accessory Factor Two in the Regulation of Phosphoenolpyruvate Carboxykinase (GTP) Gene Transcription during Diabetes* , 2001, The Journal of Biological Chemistry.

[10]  H. Shimano,et al.  Promoter Analysis of the Mouse Sterol Regulatory Element-binding Protein-1c Gene* , 2000, The Journal of Biological Chemistry.

[11]  T. Osborne,et al.  Nutrient regulation of gene expression by the sterol regulatory element binding proteins: increased recruitment of gene-specific coregulatory factors and selective hyperacetylation of histone H3 in vivo. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[12]  M. Foretz,et al.  Sterol regulatory element binding protein-1c is a major mediator of insulin action on the hepatic expression of glucokinase and lipogenesis-related genes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[13]  M. Brown,et al.  A proteolytic pathway that controls the cholesterol content of membranes, cells, and blood. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[14]  P. Farnham,et al.  c-Myc target gene specificity is determined by a post-DNAbinding mechanism. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[15]  R. Tjian,et al.  Chromatin, TAFs, and a novel multiprotein coactivator are required for synergistic activation by Sp1 and SREBP-1a in vitro. , 1998, Genes & development.

[16]  P. Edwards,et al.  CBP Is Required for Sterol-regulated and Sterol Regulatory Element-binding Protein-regulated Transcription* , 1998, The Journal of Biological Chemistry.

[17]  I. Shimomura,et al.  Regulation of sterol regulatory element binding proteins in livers of fasted and refed mice. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[18]  T. Osborne,et al.  Sterol regulation of acetyl coenzyme A carboxylase promoter requires two interdependent binding sites for sterol regulatory element binding proteins. , 1997, Journal of lipid research.

[19]  J. Goldstein,et al.  The SREBP Pathway: Regulation of Cholesterol Metabolism by Proteolysis of a Membrane-Bound Transcription Factor , 1997, Cell.

[20]  R. Roeder,et al.  Poly(ADP-ribose) polymerase enhances activator-dependent transcription in vitro. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[21]  R. Hammer,et al.  Isoform 1c of sterol regulatory element binding protein is less active than isoform 1a in livers of transgenic mice and in cultured cells. , 1997, The Journal of clinical investigation.

[22]  I. Shimomura,et al.  Differential expression of exons 1a and 1c in mRNAs for sterol regulatory element binding protein-1 in human and mouse organs and cultured cells. , 1997, The Journal of clinical investigation.

[23]  T. Osborne,et al.  Two Tandem Binding Sites for Sterol Regulatory Element Binding Proteins Are Required for Sterol Regulation of Fatty-acid Synthase Promoter* , 1996, The Journal of Biological Chemistry.

[24]  R. Tjian,et al.  SREBP transcriptional activity is mediated through an interaction with the CREB-binding protein. , 1996, Genes & development.

[25]  J. Rosenfeld,et al.  A Direct Role for Sterol Regulatory Element Binding Protein in Activation of 3-Hydroxy-3-methylglutaryl Coenzyme A Reductase Gene (*) , 1996, The Journal of Biological Chemistry.

[26]  B. Spiegelman,et al.  Dual DNA binding specificity of ADD1/SREBP1 controlled by a single amino acid in the basic helix-loop-helix domain , 1995, Molecular and cellular biology.

[27]  T. Osborne,et al.  Cooperation by Sterol Regulatory Element-binding Protein and Sp1 in Sterol Regulation of Low Density Lipoprotein Receptor Gene (*) , 1995, The Journal of Biological Chemistry.

[28]  P. Lieberman.  Identification of functional targets of the Zta transcriptional activator by formation of stable preinitiation complex intermediates , 1994, Molecular and cellular biology.

[29]  M. Kretzschmar,et al.  A novel mediator of class II gene transcription with homology to viral immediate-early transcriptional regulators , 1994, Cell.

[30]  M. Brown,et al.  Nuclear protein that binds sterol regulatory element of low density lipoprotein receptor promoter. I. Identification of the protein and delineation of its target nucleotide sequence. , 1993, The Journal of biological chemistry.

[31]  X. Wang,et al.  Nuclear protein that binds sterol regulatory element of low density lipoprotein receptor promoter. II. Purification and characterization. , 1993, The Journal of biological chemistry.

[32]  J. Friedman,et al.  Glucocorticoids regulate the induction of phosphoenolpyruvate carboxykinase (GTP) gene transcription during diabetes. , 1993, The Journal of biological chemistry.

[33]  J. Liu,et al.  Opposing actions of Fos and Jun on transcription of the phosphoenolpyruvate carboxykinase (GTP) gene. Dominant negative regulation by Fos. , 1992, The Journal of biological chemistry.

[34]  J. Liu,et al.  Identification of a thyroid hormone response element in the phosphoenolpyruvate carboxykinase (GTP) gene. Evidence for synergistic interaction between thyroid hormone and cAMP cis-regulatory elements. , 1991, The Journal of biological chemistry.

[35]  R. Roeder,et al.  Family of proteins that interact with TFIID and regulate promoter activity , 1991, Cell.

[36]  R. Roeder,et al.  Activation of class II gene transcription by regulatory factors is potentiated by a novel activity , 1991, Cell.

[37]  R. O’Brien,et al.  Regulation of phosphoenolpyruvate carboxykinase gene expression by insulin. Use of the stable transfection approach to locate an insulin responsive sequence. , 1990, Molecular endocrinology.

[38]  R. O’Brien,et al.  Identification of a sequence in the PEPCK gene that mediates a negative effect of insulin on transcription. , 1990, Science.

[39]  J. de Vente,et al.  Tissue-specific expression and dietary regulation of a chimeric phosphoenolpyruvate carboxykinase/bovine growth hormone gene in transgenic mice. , 1988, The Journal of biological chemistry.

[40]  T. Südhof,et al.  Sterol-dependent repression of low density lipoprotein receptor promoter mediated by 16-base pair sequence adjacent to binding site for transcription factor Sp1. , 1988, The Journal of biological chemistry.

[41]  W. Lamers,et al.  cAMP stimulates transcription of the gene for cytosolic phosphoenolpyruvate carboxykinase in rat liver nuclei. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[42]  D. Friend,et al.  HIGH-YIELD PREPARATION OF ISOLATED RAT LIVER PARENCHYMAL CELLS , 1969, The Journal of cell biology.

[43]  H. Leffert,et al.  [47] Liver cells , 1979 .