Transcriptional regulation of genes encoding glycolytic enzymes by hypoxia-inducible factor 1.
23 Sep 1994-Journal of Biological Chemistry (American Society for Biochemistry and Molecular Biology)-Vol. 269, Iss: 38, pp 23757-23763
TL;DR: In this article, the role of HIF-1 as a mediator of adaptive responses to hypoxia that underlie cellular and systemic oxygen homeostasis was investigated in Hep3B cells.
About: This article is published in Journal of Biological Chemistry.The article was published on 1994-09-23 and is currently open access. It has received 1817 citations till now. The article focuses on the topics: Phosphoglycerate kinase 1 & Lactate dehydrogenase A.
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TL;DR: Those Akt substrates that are most likely to contribute to the diverse cellular roles of Akt, which include cell survival, growth, proliferation, angiogenesis, metabolism, and migration are discussed.
5,505 citations
Cites background from "Transcriptional regulation of genes..."
...The frequent occurrence of PI3K-Akt pathway activation and HIFα accumulation in human cancers is likely to explain the high levels of Glut1 and enhanced glucose uptake observed in tumors (reviewed in Majumder and Sellers, 2005; Semenza, 2003)....
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...Activation of mTORC1, through Akt-mediated phosphorylation of TSC2 and PRAS40, can contribute to both HIFαdependent transcription of the Glut1 gene and capdependent translation of Glut1 mRNA (e.g., Taha et al., 1999; Zelzer et al., 1998)....
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...HIFα activation in endothelial cells and other cells leads to expression and subsequent secretion of VEGF and other angiogenic factors, thereby stimulating angiogenesis through both autocrine and paracrine signaling....
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...Akt signaling also leads to an increased production of the hypoxiainducible factor α (HIF1α and HIF2α) transcription factors, at least in part, through mTORC1-dependent translation (reviewed in Gordan and Simon, 2007; Semenza, 2003)....
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...Akt’s ability to enhance the rate of glycolysis is due, at least in part, to its ability to promote the expression of glycolytic enzymes through HIFα (e.g., Lum et al., 2007; Majumder et al., 2004; Semenza et al., 1994)....
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TL;DR: HIF-1 is implicate in the activation of VEGF transcription in hypoxic cells and this work demonstrates the involvement of Hif-1 in theactivation of V EGF transcription.
Abstract: Expression of vascular endothelial growth factor (VEGF) is induced in cells exposed to hypoxia or ischemia. Neovascularization stimulated by VEGF occurs in several important clinical contexts, including myocardial ischemia, retinal disease, and tumor growth. Hypoxia-inducible factor 1 (HIF-1) is a heterodimeric basic helix-loop-helix protein that activates transcription of the human erythropoietin gene in hypoxic cells. Here we demonstrate the involvement of HIF-1 in the activation of VEGF transcription. VEGF 5'-flanking sequences mediated transcriptional activation of reporter gene expression in hypoxic Hep3B cells. A 47-bp sequence located 985 to 939 bp 5' to the VEGF transcription initiation site mediated hypoxia-inducible reporter gene expression directed by a simian virus 40 promoter element that was otherwise minimally responsive to hypoxia. When reporters containing VEGF sequences, in the context of the native VEGF or heterologous simian virus 40 promoter, were cotransfected with expression vectors encoding HIF-1alpha and HIF-1beta (ARNT [aryl hydrocarbon receptor nuclear translocator]), reporter gene transcription was much greater in both hypoxic and nonhypoxic cells than in cells transfected with the reporter alone. A HIF-1 binding site was demonstrated in the 47-bp hypoxia response element, and a 3-bp substitution eliminated the ability of the element to bind HIF-1 and to activate transcription in response to hypoxia and/or recombinant HIF-1. Cotransfection of cells with an expression vector encoding a dominant negative form of HIF-1alpha inhibited the activation of reporter transcription in hypoxic cells in a dose-dependent manner. VEGF mRNA was not induced by hypoxia in mutant cells that do not express the HIF-1beta (ARNT) subunit. These findings implicate HIF-1 in the activation of VEGF transcription in hypoxic cells.
3,709 citations
Cites background or methods from "Transcriptional regulation of genes..."
...Critical cis-acting DNA sequences that contain essential HIF-1 binding sites have been identified in other hypoxia-inducible genes, including those encoding heme oxygenase 1 (25), inducible nitric oxide synthase (31, 35), glucose transporter 1 (7), and the glycolytic enzymes aldolase A, enolase 1, lactate dehydrogenase A, and phosphoglycerate kinase 1 (11, 12, 47, 49)....
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...Inspection of VEGF 59-flanking sequences revealed two potential HIF-1 binding sites which differed by no more than one nucleotide from the consensus binding-site sequence 59-BAC GTGCK-39 that was derived from bona fide HIF-1 binding sites previously identified in genes encoding EPO and glycolytic enzymes (49)....
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...Inclusion of the 2975 HIF-1 site, 59-TACGTGGG-39, with the previously established HIF-1 binding-site sequences (49) broadens the consensus sequence to 59-BACGTGSK-39....
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...In addition to the HIF-1 binding site at 2975, we identified a second HIF-1 site at 2306 which matched the HIF-1 binding site consensus sequence (49)....
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TL;DR: This review examines the idea that several core fluxes, including aerobic glycolysis, de novo lipid biosynthesis, and glutamine-dependent anaplerosis, form a stereotyped platform supporting proliferation of diverse cell types and regulates regulation of these fluxes by cellular mediators of signal transduction and gene expression.
3,526 citations
Cites background from "Transcriptional regulation of genes..."
...HIF-1’s targets include genes encoding glucose transporters, glycolytic enzymes, and LDH-A (O’Rourke et al., 1996; Semenza et al., 1994)....
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TL;DR: It is demonstrated that HIF-1alpha is a master regulator of cellular and developmental O2 homeostasis in Hif1a-/- embryos that manifested neural tube defects, cardiovascular malformations, and marked cell death within the cephalic mesenchyme.
Abstract: Hypoxia is an essential developmental and physiological stimulus that plays a key role in the pathophysiology of cancer, heart attack, stroke, and other major causes of mortality. Hypoxia-inducible factor 1 (HIF-1) is the only known mammalian transcription factor expressed uniquely in response to physiologically relevant levels of hypoxia. We now report that in Hif1a-/- embryonic stem cells that did not express the O2-regulated HIF-1alpha subunit, levels of mRNAs encoding glucose transporters and glycolytic enzymes were reduced, and cellular proliferation was impaired. Vascular endothelial growth factor mRNA expression was also markedly decreased in hypoxic Hif1a-/- embryonic stem cells and cystic embryoid bodies. Complete deficiency of HIF-1alpha resulted in developmental arrest and lethality by E11 of Hif1a-/- embryos that manifested neural tube defects, cardiovascular malformations, and marked cell death within the cephalic mesenchyme. In Hif1a+/+ embryos, HIF-1alpha expression increased between E8.5 and E9.5, coincident with the onset of developmental defects and cell death in Hif1a-/- embryos. These results demonstrate that HIF-1alpha is a master regulator of cellular and developmental O2 homeostasis.
2,418 citations
TL;DR: Otto Warburg's observations are re-examine in relation to the current concepts of cancer metabolism as being intimately linked to alterations of mitochondrial DNA, oncogenes and tumour suppressors, and thus readily exploitable for cancer therapy.
Abstract: Otto Warburg pioneered quantitative investigations of cancer cell metabolism, as well as photosynthesis and respiration. Warburg and co-workers showed in the 1920s that, under aerobic conditions, tumour tissues metabolize approximately tenfold more glucose to lactate in a given time than normal tissues, a phenomenon known as the Warburg effect. However, this increase in aerobic glycolysis in cancer cells is often erroneously thought to occur instead of mitochondrial respiration and has been misinterpreted as evidence for damage to respiration instead of damage to the regulation of glycolysis. In fact, many cancers exhibit the Warburg effect while retaining mitochondrial respiration. We re-examine Warburg's observations in relation to the current concepts of cancer metabolism as being intimately linked to alterations of mitochondrial DNA, oncogenes and tumour suppressors, and thus readily exploitable for cancer therapy.
2,312 citations
Cites background from "Transcriptional regulation of genes..."
...Specifically, HIF1, a heterodimer comprising HIF1α and HIF1β (also known as ARNT), activates genes that are involved in glycolysis, such as lactate dehydrogenase A ( LDHA ), the product of which contributes to a crucial component of the Warburg effect: the conversion of pyruvate to lactat...
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References
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15 Jan 2001
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Abstract: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential. Molecular Cloning, Fourth Edition, by the celebrated founding author Joe Sambrook and new co-author, the distinguished HHMI investigator Michael Green, preserves the highly praised detail and clarity of previous editions and includes specific chapters and protocols commissioned for the book from expert practitioners at Yale, U Mass, Rockefeller University, Texas Tech, Cold Spring Harbor Laboratory, Washington University, and other leading institutions. The theoretical and historical underpinnings of techniques are prominent features of the presentation throughout, information that does much to help trouble-shoot experimental problems. For the fourth edition of this classic work, the content has been entirely recast to include nucleic-acid based methods selected as the most widely used and valuable in molecular and cellular biology laboratories. Core chapters from the third edition have been revised to feature current strategies and approaches to the preparation and cloning of nucleic acids, gene transfer, and expression analysis. They are augmented by 12 new chapters which show how DNA, RNA, and proteins should be prepared, evaluated, and manipulated, and how data generation and analysis can be handled. The new content includes methods for studying interactions between cellular components, such as microarrays, next-generation sequencing technologies, RNA interference, and epigenetic analysis using DNA methylation techniques and chromatin immunoprecipitation. To make sense of the wealth of data produced by these techniques, a bioinformatics chapter describes the use of analytical tools for comparing sequences of genes and proteins and identifying common expression patterns among sets of genes. Building on thirty years of trust, reliability, and authority, the fourth edition of Mol
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TL;DR: A functionally tripartite, 50-nt hypoxia-inducible enhancer which binds several nuclear factors, one of which is induced by Hypoxia via de novo protein synthesis.
Abstract: We have identified a 50-nucleotide enhancer from the human erythropoietin gene 3'-flanking sequence which can mediate a sevenfold transcriptional induction in response to hypoxia when cloned 3' to a simian virus 40 promoter-chloramphenicol acetyltransferase reporter gene and transiently expressed in Hep3B cells Nucleotides (nt) 1 to 33 of this sequence mediate sevenfold induction of reporter gene expression when present in two tandem copies compared with threefold induction when present in a single copy, suggesting that nt 34 to 50 bind a factor which amplifies the induction signal DNase I footprinting demonstrated binding of a constitutive nuclear factor to nt 26 to 48 Mutagenesis studies revealed that nt 4 to 12 and 19 to 23 are essential for induction, as substitutions at either site eliminated hypoxia-induced expression Electrophoretic mobility shift assays identified a nuclear factor which bound to a probe spanning nt 1 to 18 but not to a probe containing a mutation which eliminated enhancer function Factor binding was induced by hypoxia, and its induction was sensitive to cycloheximide treatment We have thus defined a functionally tripartite, 50-nt hypoxia-inducible enhancer which binds several nuclear factors, one of which is induced by hypoxia via de novo protein synthesis
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TL;DR: It is demonstrated that Hif-1 DNA binding activity is also induced by hypoxia in a variety of mammalian cell lines in which the EPO gene is not transcribed, providing evidence that HIF-1 and its recognition sequence are common components of a general mammalian cellular response to Hypoxia.
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