There is controversy whether or not upstream stimulatory factors (USF) regulate the glucose responsiveness of L-pyruvate kinase (L-PK) promoter activity in hepatocytes. It has been suggested that USF-2 is required for glucose stimulation of L-PK promoter activity in single islet β-cells and INS-1 cells (Kennedy, H. J., Viollet, B., Rafiq, I., Kahn, A., and Rutter, G. A. (1997) J. Biol. Chem. 272, 20636–20640). In the present study, the tet-on system has been employed to achieve tightly controlled and inducible expression of USF-1 and -2 and their dominant-negative mutants DN-USF-1 (ΔbTDU1) and -2 (TDU2) in INS-1 cells. Quantitative Northern blot analysis shows that neither basal level nor glucose responsiveness of endogenous L-PK mRNA is affected by overexpression of USF-1 and -2. Likewise, the L-PK expression is unaltered by dominant-negative suppression of USF function. Western blotting demonstrates that USF-1 and -2 and DN-USF-1 and -2 proteins are stably expressed in nuclear fractions of INS-1 cells. Immunofluorescence staining indicates the uniform induction of these transgene-encoded proteins in the cell nuclei. Electrophoretic mobility shift assays using the L-PK promoter segment reveal that induction of USF-1 and -2 dramatically enhances the USF binding activity, whereas DN-USF-1 and -2 abolish binding. DN-USF-1 and -2 exert their dominant-negative effect by forming non-functional heterodimers with endogenous USF proteins. Carbohydrate response element-binding protein (ChREBP) was recently shown to regulate the glucose responsiveness of the L-PK promoter activity in hepatocytes. We now report the presence of this transcription factor in rat islets and INS-1 cells. Glucose stimulates ChREBP transcription in INS-1 cells, as shown by nuclear run-on experiments. Overexpression of ChREBP in INS-1 cells using the tet-on system results in a left shift of glucose responsiveness of L-PK expression and an enhanced L-PK promoter activity. Both endogenous and doxycycline-induced ChREBP proteins bind to the L-PK promoter in a glucose-dependent manner. These unprecedented results suggest that ChREBP rather than USF mediates glucose-promoted L-PK expression in insulin-secreting cells. There is controversy whether or not upstream stimulatory factors (USF) regulate the glucose responsiveness of L-pyruvate kinase (L-PK) promoter activity in hepatocytes. It has been suggested that USF-2 is required for glucose stimulation of L-PK promoter activity in single islet β-cells and INS-1 cells (Kennedy, H. J., Viollet, B., Rafiq, I., Kahn, A., and Rutter, G. A. (1997) J. Biol. Chem. 272, 20636–20640). In the present study, the tet-on system has been employed to achieve tightly controlled and inducible expression of USF-1 and -2 and their dominant-negative mutants DN-USF-1 (ΔbTDU1) and -2 (TDU2) in INS-1 cells. Quantitative Northern blot analysis shows that neither basal level nor glucose responsiveness of endogenous L-PK mRNA is affected by overexpression of USF-1 and -2. Likewise, the L-PK expression is unaltered by dominant-negative suppression of USF function. Western blotting demonstrates that USF-1 and -2 and DN-USF-1 and -2 proteins are stably expressed in nuclear fractions of INS-1 cells. Immunofluorescence staining indicates the uniform induction of these transgene-encoded proteins in the cell nuclei. Electrophoretic mobility shift assays using the L-PK promoter segment reveal that induction of USF-1 and -2 dramatically enhances the USF binding activity, whereas DN-USF-1 and -2 abolish binding. DN-USF-1 and -2 exert their dominant-negative effect by forming non-functional heterodimers with endogenous USF proteins. Carbohydrate response element-binding protein (ChREBP) was recently shown to regulate the glucose responsiveness of the L-PK promoter activity in hepatocytes. We now report the presence of this transcription factor in rat islets and INS-1 cells. Glucose stimulates ChREBP transcription in INS-1 cells, as shown by nuclear run-on experiments. Overexpression of ChREBP in INS-1 cells using the tet-on system results in a left shift of glucose responsiveness of L-PK expression and an enhanced L-PK promoter activity. Both endogenous and doxycycline-induced ChREBP proteins bind to the L-PK promoter in a glucose-dependent manner. These unprecedented results suggest that ChREBP rather than USF mediates glucose-promoted L-PK expression in insulin-secreting cells. liver-type pyruvate kinase basic/helix-loop-helix/leucine zipper upstream stimulatory factor electrophoretic mobility shift assay carbohydrate response element ChoRE-binding protein dominant-negative mutant of USF A set of glucose-responsive genes in hepatocytes has evolved to control the conversion of glucose into triglycerides, when food is sufficient, for storing energy to be used during fasting (1Towle H.C. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13476-13478Crossref PubMed Scopus (37) Google Scholar). The pancreatic β-cell responds to increased glucose metabolism by releasing insulin, which is essential for maintaining glucose and fatty acid homeostasis (1Towle H.C. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13476-13478Crossref PubMed Scopus (37) Google Scholar). L-pyruvate kinase (L-PK)1 represents a typical glucose-responsive gene in both hepatocytes and pancreatic β-cells (2Marie S. Diaz-Guerra M.J. Miquerol L. Kahn A. Iynedjian P.B. J. Biol. Chem. 1993; 268: 23881-23890Abstract Full Text PDF PubMed Google Scholar, 3Lefrancois-Martinez A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar, 5Kennedy H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar, 6Vallet V.S. Henrion A.A. Bucchini D. Casado M. Raymondjean M. Kahn A. Vaulont S. J. Biol. Chem. 1997; 272: 21944-21949Abstract Full Text Full Text PDF PubMed Scopus (94) Google Scholar, 7Kaytor E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google Scholar, 8Kawaguchi T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google Scholar, 9Yamashita H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). is controversy the of the transcription factor the L-PK A carbohydrate response element the by has been in the L-PK promoter A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar). has been that a of the basic/helix-loop-helix/leucine zipper of transcription factors be in A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar). stimulatory factor (USF) was the transcription factor of this to regulate the glucose response of the L-PK gene promoter in hepatocytes A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar). E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google results and of USF in glucose of the L-PK gene in hepatocytes. of in in a glucose responsiveness of L-PK an for V.S. Henrion A.A. Bucchini D. Casado M. Raymondjean M. Kahn A. Vaulont S. J. Biol. Chem. 1997; 272: 21944-21949Abstract Full Text Full Text PDF PubMed Scopus (94) Google Scholar). In of into the of single INS-1 cells a of L-PK promoter activity H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar). E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google that overexpression of a dominant-negative of USF in INS-1 cells to the L-PK promoter activity and the of USF in in INS-1 cells. The present was to these We employed the tet-on system M. S. G. G. H. 1995; 268: PubMed Scopus Google in INS-1 cells to achieve tightly controlled and expression of USF-1 and -2 and their dominant-negative mutants A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google DN-USF-1 (ΔbTDU1) and -2 These cell of the of USF in the of endogenous L-PK mRNA of that a ChoRE-binding protein of to the L-PK promoter in a glucose-dependent and that of hepatocytes with ChREBP to increased L-PK promoter activity T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google Scholar, 9Yamashita H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). The present demonstrates that ChREBP is expressed in pancreatic islets and in INS-1 cells. We the that ChREBP left the glucose responsiveness of endogenous L-PK expression in INS-1 cells for inducible ChREBP is controversy the of USF in the of L-PK expression in pancreatic and INS-1 cells A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar, 5Kennedy H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar, 6Vallet V.S. Henrion A.A. Bucchini D. Casado M. Raymondjean M. Kahn A. Vaulont S. J. Biol. Chem. 1997; 272: 21944-21949Abstract Full Text Full Text PDF PubMed Scopus (94) Google Scholar, 7Kaytor E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google Scholar). results of the L-PK activity using of hepatocytes with USF and A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar). the expression of USF and not in V.S. Henrion A.A. Bucchini D. Casado M. Raymondjean M. Kahn A. Vaulont S. J. Biol. Chem. 1997; 272: 21944-21949Abstract Full Text Full Text PDF PubMed Scopus (94) Google a response of L-PK expression in the of be to in expression of genes as the USF the expression of this gene P.B. Biochem. J. PubMed Scopus Google Scholar). of USF into the of INS-1 cells the L-PK promoter activity H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google and activity be with this H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google is by E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google suggest that expression of not with L-PK promoter activity. the expression level and of in the was not E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google present was to the of USF in the of L-PK expression in a controlled manner. We inducible expression of USF-1 and -2 and DN-USF-1 and The expression level and of these transgene-encoded proteins are in We that these nuclear USF and proteins are in a manner. that induction of USF-1 and -2 to an in the USF binding to the L-PK whereas induction of DN-USF-1 and -2 endogenous USF binding activity. with that DN-USF-1 dominant-negative by forming non-functional heterodimers with endogenous to for the binding. using Northern blot analysis the of and of USF glucose responsiveness of endogenous L-PK We that USF has effect basal or L-PK expression in INS-1 cells. induction of or not the L-PK promoter activity in INS-1 cells, which the of E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google the report of H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google has been suggested that a transcription factor USF the glucose response of the L-PK promoter Towle H.C. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google this protein to bind to the L-PK and as ChREBP by the In ChREBP to the L-PK promoter and to the of hepatocytes in a glucose-dependent T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google Scholar, 9Yamashita H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). overexpression of ChREBP in hepatocytes by enhanced L-PK promoter activity T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). The present the We that ChREBP is expressed in rat islets and INS-1 cells. that glucose stimulates the expression of ChREBP the level in INS-1 cells. results the that ChREBP the expression of endogenous L-PK mRNA in insulin-secreting cells by the L-PK promoter activity. in INS-1 cells both endogenous and doxycycline-induced ChREBP proteins bind to the L-PK promoter in response to It has been in hepatocytes that glucose the nuclear of ChREBP protein by of in the and stimulates the binding activity by of in the T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google Scholar, 9Yamashita H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). It to be whether to INS-1 that ChREBP rather than USF the glucose responsiveness of L-PK expression in INS-1 cells. It is that a in the of genes in hepatocytes and A set of glucose-responsive genes in hepatocytes has evolved to control the conversion of glucose into triglycerides, when food is sufficient, for storing energy to be used during fasting (1Towle H.C. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13476-13478Crossref PubMed Scopus (37) Google Scholar). The pancreatic β-cell responds to increased glucose metabolism by releasing insulin, which is essential for maintaining glucose and fatty acid homeostasis (1Towle H.C. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13476-13478Crossref PubMed Scopus (37) Google Scholar). L-pyruvate kinase (L-PK)1 represents a typical glucose-responsive gene in both hepatocytes and pancreatic β-cells (2Marie S. Diaz-Guerra M.J. Miquerol L. Kahn A. Iynedjian P.B. J. Biol. Chem. 1993; 268: 23881-23890Abstract Full Text PDF PubMed Google Scholar, 3Lefrancois-Martinez A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar, 5Kennedy H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar, 6Vallet V.S. Henrion A.A. Bucchini D. Casado M. Raymondjean M. Kahn A. Vaulont S. J. Biol. Chem. 1997; 272: 21944-21949Abstract Full Text Full Text PDF PubMed Scopus (94) Google Scholar, 7Kaytor E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google Scholar, 8Kawaguchi T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google Scholar, 9Yamashita H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). is controversy the of the transcription factor the L-PK A carbohydrate response element the by has been in the L-PK promoter A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar). has been that a of the basic/helix-loop-helix/leucine zipper of transcription factors be in A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar). stimulatory factor (USF) was the transcription factor of this to regulate the glucose response of the L-PK gene promoter in hepatocytes A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar). E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google results and of USF in glucose of the L-PK gene in hepatocytes. of in in a glucose responsiveness of L-PK an for V.S. Henrion A.A. Bucchini D. Casado M. Raymondjean M. Kahn A. Vaulont S. J. Biol. Chem. 1997; 272: 21944-21949Abstract Full Text Full Text PDF PubMed Scopus (94) Google Scholar). In of into the of single INS-1 cells a of L-PK promoter activity H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar). E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google that overexpression of a dominant-negative of USF in INS-1 cells to the L-PK promoter activity and the of USF in in INS-1 cells. The present was to these We employed the tet-on system M. S. G. G. H. 1995; 268: PubMed Scopus Google in INS-1 cells to achieve tightly controlled and expression of USF-1 and -2 and their dominant-negative mutants A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google DN-USF-1 (ΔbTDU1) and -2 These cell of the of USF in the of endogenous L-PK mRNA of that a ChoRE-binding protein of to the L-PK promoter in a glucose-dependent and that of hepatocytes with ChREBP to increased L-PK promoter activity T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google Scholar, 9Yamashita H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). The present demonstrates that ChREBP is expressed in pancreatic islets and in INS-1 cells. We the that ChREBP left the glucose responsiveness of endogenous L-PK expression in INS-1 cells for inducible ChREBP is controversy the of USF in the of L-PK expression in pancreatic and INS-1 cells A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar, 5Kennedy H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar, 6Vallet V.S. Henrion A.A. Bucchini D. Casado M. Raymondjean M. Kahn A. Vaulont S. J. Biol. Chem. 1997; 272: 21944-21949Abstract Full Text Full Text PDF PubMed Scopus (94) Google Scholar, 7Kaytor E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google Scholar). results of the L-PK activity using of hepatocytes with USF and A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar). the expression of USF and not in V.S. Henrion A.A. Bucchini D. Casado M. Raymondjean M. Kahn A. Vaulont S. J. Biol. Chem. 1997; 272: 21944-21949Abstract Full Text Full Text PDF PubMed Scopus (94) Google a response of L-PK expression in the of be to in expression of genes as the USF the expression of this gene P.B. Biochem. J. PubMed Scopus Google Scholar). of USF into the of INS-1 cells the L-PK promoter activity H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google and activity be with this H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google is by E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google suggest that expression of not with L-PK promoter activity. the expression level and of in the was not E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google present was to the of USF in the of L-PK expression in a controlled manner. We inducible expression of USF-1 and -2 and DN-USF-1 and The expression level and of these transgene-encoded proteins are in We that these nuclear USF and proteins are in a manner. that induction of USF-1 and -2 to an in the USF binding to the L-PK whereas induction of DN-USF-1 and -2 endogenous USF binding activity. with that DN-USF-1 dominant-negative by forming non-functional heterodimers with endogenous to for the binding. using Northern blot analysis the of and of USF glucose responsiveness of endogenous L-PK We that USF has effect basal or L-PK expression in INS-1 cells. induction of or not the L-PK promoter activity in INS-1 cells, which the of E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google the report of H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google has been suggested that a transcription factor USF the glucose response of the L-PK promoter Towle H.C. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google this protein to bind to the L-PK and as ChREBP by the In ChREBP to the L-PK promoter and to the of hepatocytes in a glucose-dependent T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google Scholar, 9Yamashita H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). overexpression of ChREBP in hepatocytes by enhanced L-PK promoter activity T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). The present the We that ChREBP is expressed in rat islets and INS-1 cells. that glucose stimulates the expression of ChREBP the level in INS-1 cells. results the that ChREBP the expression of endogenous L-PK mRNA in insulin-secreting cells by the L-PK promoter activity. in INS-1 cells both endogenous and doxycycline-induced ChREBP proteins bind to the L-PK promoter in response to It has been in hepatocytes that glucose the nuclear of ChREBP protein by of in the and stimulates the binding activity by of in the T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google Scholar, 9Yamashita H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). It to be whether to INS-1 that ChREBP rather than USF the glucose responsiveness of L-PK expression in INS-1 cells. It is that a in the of genes in hepatocytes and There is controversy the of USF in the of L-PK expression in pancreatic and INS-1 cells A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar, 5Kennedy H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar, 6Vallet V.S. Henrion A.A. Bucchini D. Casado M. Raymondjean M. Kahn A. Vaulont S. J. Biol. Chem. 1997; 272: 21944-21949Abstract Full Text Full Text PDF PubMed Scopus (94) Google Scholar, 7Kaytor E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google Scholar). results of the L-PK activity using of hepatocytes with USF and A.M. Martinez A. Antoine B. Raymondjean M. Kahn A. J. Biol. Chem. 1995; 270: 2640-2643Abstract Full Text Full Text PDF PubMed Scopus (112) Google Scholar, 4Kaytor E.N. Shih H. Towle H.C. J. Biol. Chem. 1997; 272: 7525-7531Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar). the expression of USF and not in V.S. Henrion A.A. Bucchini D. Casado M. Raymondjean M. Kahn A. Vaulont S. J. Biol. Chem. 1997; 272: 21944-21949Abstract Full Text Full Text PDF PubMed Scopus (94) Google a response of L-PK expression in the of be to in expression of genes as the USF the expression of this gene P.B. Biochem. J. PubMed Scopus Google Scholar). of USF into the of INS-1 cells the L-PK promoter activity H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google and activity be with this H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google is by E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google suggest that expression of not with L-PK promoter activity. the expression level and of in the was not E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google Scholar). The present was to the of USF in the of L-PK expression in a controlled manner. We inducible expression of USF-1 and -2 and DN-USF-1 and The expression level and of these transgene-encoded proteins are in We that these nuclear USF and proteins are in a manner. that induction of USF-1 and -2 to an in the USF binding to the L-PK whereas induction of DN-USF-1 and -2 endogenous USF binding activity. with that DN-USF-1 dominant-negative by forming non-functional heterodimers with endogenous to for the binding. using Northern blot analysis the of and of USF glucose responsiveness of endogenous L-PK We that USF has effect basal or L-PK expression in INS-1 cells. induction of or not the L-PK promoter activity in INS-1 cells, which the of E.N. Qian J. Towle H.C. Olson L.K. Mol Cell Biochem. 2000; 210: 13-21Crossref PubMed Google the report of H.J. Viollet B. Rafiq I. Kahn A. Rutter G.A. J. Biol. Chem. 1997; 272: 20636-20640Abstract Full Text Full Text PDF PubMed Scopus (65) Google Scholar). It has been suggested that a transcription factor USF the glucose response of the L-PK promoter Towle H.C. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google this protein to bind to the L-PK and as ChREBP by the In ChREBP to the L-PK promoter and to the of hepatocytes in a glucose-dependent T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google Scholar, 9Yamashita H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). overexpression of ChREBP in hepatocytes by enhanced L-PK promoter activity T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). The present the We that ChREBP is expressed in rat islets and INS-1 cells. that glucose stimulates the expression of ChREBP the level in INS-1 cells. results the that ChREBP the expression of endogenous L-PK mRNA in insulin-secreting cells by the L-PK promoter activity. in INS-1 cells both endogenous and doxycycline-induced ChREBP proteins bind to the L-PK promoter in response to It has been in hepatocytes that glucose the nuclear of ChREBP protein by of in the and stimulates the binding activity by of in the T. Takenoshita M. Kabashima T. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 13710-13715Crossref PubMed Scopus (298) Google Scholar, 9Yamashita H. Takenoshita M. Sakurai M. Bruick R.K. Henzel D. Uyeda K. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: PubMed Scopus Google Scholar). It to be whether to INS-1 cells. We that ChREBP rather than USF the glucose responsiveness of L-PK expression in INS-1 cells. It is that a in the of genes in hepatocytes and We are to D. and for We are to A. Kahn and -2 and USF M. and -2 and USF H. Towle B. Iynedjian H. and
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