Regulation of human heme oxygenase-1 gene expression under thermal stress

Heme oxygenase-1 is an essential enzyme in heme catabolism, and its human gene promoter contains a putative heat shock element (HHO-HSE). This study was designed to analyze the regulation of human heme oxygenase-1 gene expression under thermal stress. The amounts of heme oxygenase-1 protein were not increased by heat shock (incubation at 42 degrees C) in human alveolar macrophages and in a human erythroblastic cell line, YN-1–0-A, whereas heat shock protein 70 (HSP70) was noticeably induced. However, heat shock factor does bind in vitro to HHO-HSE and the synthetic HHO-HSE by itself is sufficient to confer the increase in the transient expression of a reporter gene upon heat shock. The deletion of the sequence, located downstream from HHO-HSE, resulted in the activation of a reporter gene by heat shock. These results suggest that HHO-HSE is potentially functional but is repressed in vivo. Interestingly, heat shock abolished the remarkable increase in the levels of heme oxygenase-1 mRNA in YN-1–0-A cells treated with hemin or cadmium, in which HSP70 mRNA was noticeably induced. Furthermore, transient expression assays showed that heat shock inhibits the cadmium-mediated activation of the heme oxygenase-1 promoter, whereas the HSP70 gene promoter was activated upon heat shock. Such regulation of heme oxygenase-1 under thermal stress may be of physiologic significance in erythroid cells.

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Regulation of Heme Oxygenase-1 Gene Expression by Anoxia and Reoxygenation in Primary Rat Hepatocyte Cultures

Experimental Biology and Medicine ◽

10.1177/15353702-0322805-51 ◽

2003 ◽

Vol 228 (5) ◽

pp. 584-589 ◽

Cited By ~ 15

Author(s):

Andreas Ohlmann ◽

Susanne Giffhorn-Katz ◽

Ivonne Becker ◽

Norbert Katz ◽

Stephan Immenschuh

Keyword(s):

Gene Expression ◽

Mrna Expression ◽

Heme Oxygenase ◽

Heme Oxygenase 1 ◽

Protein Synthesis Inhibitor ◽

Mrna Levels ◽

Rat Hepatocyte ◽

Hsp70 Mrna ◽

Hepatocyte Cultures ◽

Antioxidant Agents

Heme oxygenase (HO) catalyzes the rate-limiting enzymatic step of heme degradation and regulates the cellular heme content. Gene expression of the inducible isoform of HO, HO-1, is upregulated in response to various oxidative stress stimuli. To investigate the regulatory role of anoxia and reoxygenation (A/R) on hepatic HO-1 gene expression, primary cultures of rat hepatocytes were exposed after an anoxia of 4 hr to normal oxygen tension for various lengths of time. For comparison, gene expression of the noninducible HO isoform, HO-2, and that of the heat-shock protein 70 (HSP70) were determined. During reoxygenation, a marked increase of HO-1 and HSP70 steady-state mRNA levels was observed, whereas no alteration of HO-2 mRNA levels occurred. Corresponding to HO-1 mRNA, an increase of HO-1 protein expression was determined by Western blot analysis. The anoxia-dependent induction of HO-1 was prevented by pretreatment with the transcription inhibitor, actinomycin D, but not by the protein synthesis inhibitor, cycloheximide, suggesting a transcriptional regulatory mechanism. After exposure of hepatocytes to anoxia, the relative levels of oxidized glutathione increased within the first 40 min of reoxygenation. Pretreament of cell cultures with the antioxidant agents, β-carotene and allopurinol, before exposure to A/R led to a marked decrease of HO-1 and HSP70 mRNA expression during reoxygenation. An even more pronounced reduction of mRNA expression was observed after exposure to desferrioxamine. Taken together, the data demonstrate that HO-1 gene expression in rat hepatocyte cultures after A/R is upregulated by a transcriptional mechanism that may be, in part, mediated via the generation of ROS and the glutathione system.

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Induction of heme oxygenase-1 and heat shock protein 70 in rat hepatocytes: The role of calcium signaling

Cellular & Molecular Biology Letters ◽

10.2478/s11658-006-0052-0 ◽

2007 ◽

Vol 12 (1) ◽

Cited By ~ 6

Author(s):

Malte Silomon ◽

Inge Bauer ◽

Michael Bauer ◽

Julia Nolting ◽

Markus Paxian ◽

...

Keyword(s):

Gene Expression ◽

Stress Response ◽

Heat Shock ◽

Heme Oxygenase ◽

Rat Hepatocytes ◽

Glutathione Depletion ◽

Heme Oxygenase 1 ◽

Hsp70 Gene ◽

Stress Response Genes

AbstractStress response genes including heat shock proteins are induced under a variety of conditions to confer cellular protection. This study investigated the role of calcium signaling in the induction of two stress response genes, heme oxygenase-1/hsp32 and hsp70, in isolated rat hepatocytes. Both genes were induced by cellular glutathione depletion. This induction could be inhibited by BAPTA-AM. Culturing in a calcium-free medium prevented the induction of hsp70 gene expression after glutathione depletion without affecting heme oxygenase-1 gene expression. Thapsigargin increased the gene expression of heme oxygenase-1 but not that of hsp70. Thapsigargin-induced heme oxygenase-1 induction was completely inhibited by BAPTA-AM. Incubation with the Ca2+-ionophore A23187 augmented heme oxygenase-1 (two-fold) and hsp70 (5.2-fold) mRNA levels. Our data suggests a significant role of Ca2+-dependent pathways in the induction of the two stress genes. An increase in the cytoplasmic Ca2+ activity seems to play a key role in the cascade of signaling leading to the induction of the two genes. However, the source of Ca2+ that fluxes into the cytoplasm seems to be different. Our data provides evidence for a compartmentalization of calcium fluxes, i.e. the Ca2+ flux from intracellular stores (e.g. the endoplasmic reticulum) plays a major role in the induction of heme oxygenase-1. By contrast, Ca2+ flux from the extracellular medium seems to be a mechanism initiating the cellular signaling cascade leading to hsp70 gene induction.

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PS-074 Heme Oxygenase-1 Gene Promoter Polymorphism And Insulin Resistance Are Associated With Susceptibility To Non-alcoholic Fatty Liver Disease In Children

Archives of Disease in Childhood ◽

10.1136/archdischild-2014-307384.371 ◽

2014 ◽

Vol 99 (Suppl 2) ◽

pp. A138.3-A139

Author(s):

P Chang ◽

Y Lin ◽

K Liu ◽

S Yeh

Keyword(s):

Insulin Resistance ◽

Liver Disease ◽

Heme Oxygenase ◽

Fatty Liver Disease ◽

Gene Promoter ◽

Promoter Polymorphism ◽

Heme Oxygenase 1 ◽

Alcoholic Fatty Liver ◽

Alcoholic Fatty Liver Disease ◽

Gene Promoter Polymorphism

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Microsatellite polymorphism in the heme oxygenase-1 gene promoter is not associated with alcoholic liver disease severity

Hepatology ◽

10.1002/hep.26534 ◽

2013 ◽

Vol 59 (1) ◽

pp. 352-353 ◽

Cited By ~ 1

Author(s):

Anne Lemaire ◽

Eric Trépo ◽

Romy Ouziel ◽

Thierry Gustot ◽

Christophe Moreno ◽

...

Keyword(s):

Liver Disease ◽

Disease Severity ◽

Heme Oxygenase ◽

Alcoholic Liver Disease ◽

Gene Promoter ◽

Microsatellite Polymorphism ◽

Heme Oxygenase 1

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Adenovirus-mediated heme oxygenase-1 gene transfer to neonatal porcine islet-like cluster cells: the effects on gene expression and protection from cell stress

BioChip Journal ◽

10.1007/s13206-012-6108-5 ◽

2012 ◽

Vol 6 (1) ◽

pp. 56-64 ◽

Cited By ~ 1

Author(s):

Hye-Jung Yeom ◽

Han Ro ◽

Sol Ji Park ◽

Ju Ho Hong ◽

Bumrae Cho ◽

...

Keyword(s):

Gene Expression ◽

Gene Transfer ◽

Heme Oxygenase ◽

Cell Stress ◽

Heme Oxygenase 1 ◽

Porcine Islet

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Modular recognition of 5-base-pair DNA sequence motifs by human heat shock transcription factor

Molecular and Cellular Biology ◽

10.1128/mcb.11.7.3504-3514.1991 ◽

1991 ◽

Vol 11 (7) ◽

pp. 3504-3514

Author(s):

N F Cunniff ◽

J Wagner ◽

W D Morgan

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Gene Promoter ◽

Heat Shock Transcription Factor ◽

Heat Shock Gene ◽

Gene Promoters ◽

Sequence Motifs ◽

Binding Modes ◽

Heat Shock Element

We investigated the recognition of the conserved 5-bp repeated motif NGAAN, which occurs in heat shock gene promoters of Drosophila melanogaster and other eukaryotic organisms, by human heat shock transcription factor (HSF). Extended heat shock element mutants of the human HSP70 gene promoter, containing additional NGAAN blocks flanking the original element, showed significantly higher affinity than the wild-type promoter element for human HSF in vitro. Protein-DNA contact positions were identified by hydroxyl radical protection, diethyl pyrocarbonate interference, and DNase I footprinting. New contacts in the mutant HSE constructs corresponded to the locations of additional NGAAN motifs. The pattern of binding indicated the occurrence of multiple DNA binding modes for HSF with the various constructs and was consistent with an oligomeric, possibly trimeric, structure of the protein. In contrast to the improved binding, the extended heat shock element mutant constructs did not exhibit dramatically increased heat-inducible transcription in transient expression assays with HeLa cells.

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Identification of cis and trans components of a novel heat shock stress regulatory pathway in Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.13.1.248-256.1993 ◽

1993 ◽

Vol 13 (1) ◽

pp. 248-256

Author(s):

N Kobayashi ◽

K McEntee

Keyword(s):

Saccharomyces Cerevisiae ◽

Stress Response ◽

Heat Shock ◽

Reporter Gene ◽

Heat Shock Element ◽

Cis Acting ◽

Heat Shock Stress ◽

Specific Complex ◽

Cis And Trans ◽

Shock Stress

The stress-responsive DDR2 gene (previously called DDRA2) of Saccharomyces cerevisiae is transcribed at elevated levels following stress caused by heat shock or DNA damage. Previously, we identified a 51-bp promoter fragment, oligo31/32, which conferred heat shock inducibility on the heterologous CYC1-lacZ reporter gene in S. cerevisiae (N. Kobayashi and K. McEntee, Proc. Natl. Acad. Sci. USA 87:6550-6554, 1990). Using a series of synthetic oligonucleotides, we have identified a pentanucleotide, CCCCT (C4T), as an essential component of this stress response sequence. This element is not a binding site for the well-characterized heat shock transcription factor which recognizes a distinct cis-acting heat shock element in the promoters of many heat shock genes. Here we demonstrate the ability of oligonucleotides containing the C4T sequence to confer heat shock inducibility on the reporter gene and show that the presence of two such elements produces more than additive effects on induction. Gel retardation experiments have been used to demonstrate specific complex formation between C4T-containing fragments and one or more yeast proteins. Formation of these complexes was not competed by fragments containing mutations in the C4T sequence nor by heat shock element-containing competitor DNAs. Fragments containing the C4T element bound to a single 140-kDa polypeptide, distinct from heat shock transcription factors in yeast crude extracts. These experiments identify key cis- and trans-acting components of a novel heat shock stress response pathway in S. cerevisiae.

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