The Yeast Pyruvate Kinase Gene is Regulated at Multiple Levels

To clarify carbon source-dependent control of the glycolytic pathway in the yeast Saccharomyces cerevisiae, we have initiated a study of transcriptional regulation of the pyruvate kinase gene (PYK). By deletion analysis of the 5'-noncoding region of the PYK gene, we have identified an upstream activating sequence (UASPYK1) located between 634 and 653 nucleotides upstream of the initiating ATG codon. The promoter activity of the PYK 5'-noncoding region was abolished when the sequence containing the UASPYK1 was deleted from the region. Synthetic UASPYK1 (26mer), in either orientation, was able to restore the transcriptional activity of UAS-depleted mutants when placed upstream of the TATA sequence located at -199 (ATG as +1). While the UASPYK1 was required for basal to intermediate levels of transcriptional activation, a sequence between -714 and -811 was found to be necessary for full activation. On the other hand, a sequence between -344 and -468 was found to be responsible for transcriptional repression of the PYK gene when yeast cells were grown on nonfermentable carbon sources. This upstream repressible sequence also repressed transcription, although to a lesser extent, when glucose was present in the medium. The possible mechanism for carbon source-dependent regulation of PYK expression through these cis-acting regulatory elements is discussed.

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Spot 14 protein interacts and co-operates with chicken ovalbumin upstream promoter-transcription factor 1 in the transcription of the L-type pyruvate kinase gene through a specificity protein 1 (Sp1) binding site

Biochemical Journal ◽

10.1042/bj3580175 ◽

2001 ◽

Vol 358 (1) ◽

pp. 175-183 ◽

Cited By ~ 11

Author(s):

Emmanuel COMPE ◽

Georges de SOUSA ◽

Kamel FRANCÇOIS ◽

Régis ROCHE ◽

Roger RAHMANI ◽

...

Keyword(s):

Transcription Factor ◽

Pyruvate Kinase ◽

Nuclear Proteins ◽

Kinase Gene ◽

Specificity Protein 1 ◽

Upstream Promoter ◽

Spot 14 ◽

Specificity Protein ◽

Transcription Factor 1 ◽

Chicken Ovalbumin

In hepatocytes, the amount of the Spot 14 (S14) protein is closely related to the full expression of enzymes involved in the glycolytic and lipogenic pathways. In the present study we address the role played by this protein in the control of transcription of the L-type pyruvate kinase (L-PK) gene in primary hepatocytes. We show that human S14, which by itself does not bind to the L-PK promoter, physically interacts with the human chicken ovalbumin upstream promoter-transcription factor 1 (COUP-TF1) and induces the switch of this factor from a repressor to an activator. However, the enhancing activity of S14 and COUP-TF1 depends on the presence of a proximal GC-rich box (the L0 element) that specifically binds nuclear proteins from the livers of rats fed a glucose-rich diet. Moreover, the L0 element, which strongly binds dephosphorylated specificity protein 1 (Sp1), loses all affinity when this factor is phosphorylated by cAMP-dependent protein kinase. Mutations that affect binding of Sp1 and nuclear proteins to the L0 box also decrease basal transcription and impair glucose responsiveness of the promoter. These results therefore shed light on the mechanism by which the S14 protein, whose concentration rapidly rises after glucose intake, contributes to the full activity of the L-PK promoter.

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Positive and negative regulation of gene expression by insulin and glucagon: The model of L-type pyruvate kinase gene

Biochimie ◽

10.1016/0300-9084(91)90072-9 ◽

1991 ◽

Vol 73 (1) ◽

pp. 41-45 ◽

Cited By ~ 3

Author(s):

M. Raymondjean ◽

S. Voulont ◽

M. Cognet ◽

J.F. Decaux ◽

N. Puzenat ◽

...

Keyword(s):

Gene Expression ◽

Pyruvate Kinase ◽

Regulation Of Gene Expression ◽

Negative Regulation ◽

Kinase Gene

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Functional synergism in the carbohydrate-induced activation of liver-type pyruvate kinase gene expression

Biochemical Journal ◽

10.1042/bj3080105 ◽

1995 ◽

Vol 308 (1) ◽

pp. 105-111 ◽

Cited By ~ 18

Author(s):

Z Liu ◽

H C Towle

Keyword(s):

Pyruvate Kinase ◽

Binding Sites ◽

Transcriptional Level ◽

Primary Hepatocytes ◽

Kinase Gene ◽

Glucose Response ◽

Hepatic Expression ◽

Elevated Glucose ◽

Response Prompting ◽

Late Transcription

Hepatic expression of the liver-type pyruvate kinase (L-PK) gene is induced at the transcriptional level by increased carbohydrate metabolism in the rat. The carbohydrate response of the L-PK gene requires sequences from -171 to -124, which encompass adjacent major late transcription factor (MLTF)-like and hepatic nuclear factor (HNF)-4 binding sites. Neither site alone is capable of conferring a response, prompting us to explore the mechanism of synergy between the MLTF-like factor and HNF-4. Spacing requirements between the two factor binding sites were tested by generating a series of mutations that altered the distance between these sites. Surprisingly, all of the constructs with spacing mutations were capable of responding to elevated glucose when introduced into primary hepatocytes. Thus the glucose response does not depend on the rigid phasing of the MLTF-like and HNF-4 factors, suggesting that the factors binding to these two sites do not interact directly with each other. Substitution or inversion of the PK HNF-4 site abrogated the response to glucose and also significantly suppressed the promoter activity under non-inducing conditions. We conclude that the MLTF-like factor and HNF-4 co-operate functionally to maintain the basal activity, as well as the carbohydrate responsiveness, of the L-PK gene. A mechanism other than co-operative DNA binding is responsible for the synergism.

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Zinc Deficiency Changes Preferred Macronutrient Intake in Subpopulations of Sprague-Dawley Outbred Rats and Reduces Hepatic Pyruvate Kinase Gene Expression

Journal of Nutrition ◽

10.1093/jn/128.1.43 ◽

1998 ◽

Vol 128 (1) ◽

pp. 43-49 ◽

Cited By ~ 35

Author(s):

Kathleen J. Kennedy ◽

Tia M. Rains ◽

Neil F. Shay

Keyword(s):

Gene Expression ◽

Pyruvate Kinase ◽

Zinc Deficiency ◽

Macronutrient Intake ◽

Sprague Dawley ◽

Kinase Gene

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c-Myc and ChREBP regulate glucose-mediated expression of the L-type pyruvate kinase gene in INS-1-derived 832/13 cells

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00357.2006 ◽

2007 ◽

Vol 293 (1) ◽

pp. E48-E56 ◽

Cited By ~ 26

Author(s):

J. Jason Collier ◽

Pili Zhang ◽

Kim B. Pedersen ◽

Susan J. Burke ◽

John W. Haycock ◽

...

Keyword(s):

Pyruvate Kinase ◽

Β Cells ◽

Pancreatic Β Cells ◽

Heterologous Promoter ◽

Kinase Gene ◽

Glucose Response ◽

Glucose Flux ◽

Element Binding Protein ◽

Regulated Gene Expression

Increased glucose flux generates metabolic signals that control transcriptional programs through poorly understood mechanisms. Previously, we demonstrated a necessity in hepatocytes for c-Myc in the regulation of a prototypical glucose-responsive gene, L-type pyruvate kinase (L-PK) (Collier JJ, Doan TT, Daniels MC, Schurr JR, Kolls JK, Scott DK. J Biol Chem 278: 6588–6595, 2003). Pancreatic β-cells have many features in common with hepatocytes with respect to glucose-regulated gene expression, and in the present study we determined whether c-Myc was required for the L-PK glucose response in insulin-secreting (INS-1)-derived 832/13 cells. Glucose increased c-Myc abundance and association with its heterodimer partner, Max. Manipulations that prevented the formation of a functional c-Myc/Max heterodimer reduced the expression of the L-PK gene. In addition, glucose augmented the binding of carbohydrate response element binding protein (ChREBP), c-Myc, and Max to the promoter of the L-PK gene in situ. The transactivation of ChREBP, but not of c-Myc, was dependent on high glucose concentrations in the contexts of either the L-PK promoter or a heterologous promoter. The glucose-mediated transactivation of ChREBP was independent of mutations that alter phosphorylation sites thought to regulate the cellular location of ChREBP. We conclude that maximal glucose-induced expression of the L-PK gene in INS-1-derived 832/13 cells involves increased c-Myc abundance, recruitment of c-Myc, Max, and ChREBP to the promoter, and a glucose-stimulated increase in ChREBP transactivation.

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