Endoplasmic reticulum (ER) stress signal impairs erythropoietin production: a role for ATF4.

University of Tokyo School of Medicine.
AJP Cell Physiology (Impact Factor: 3.78). 12/2012; 304(4). DOI: 10.1152/ajpcell.00153.2012
Source: PubMed


Hypoxia upregulates hypoxia inducible factor (HIF) pathway and ER stress signal, unfolded protein response (UPR). The crosstalk of both signals affects the pathogenic alteration by hypoxia. Here, we showed that ER stress induced by tunicamycin or thapsigargin suppressed inducible (CoCl(2) or hypoxia) transcription of erythropoietin (EPO), a representative HIF target gene, in HepG2. This suppression was inversely correlated with UPR activation, as estimated by expression of UPR regulator GRP78, and restored by an ER stress inhibitor, salubrinal, in association with normalization of the UPR state. Importantly, the decreased EPO expression was also observed in HepG2 overexpressing UPR transcription factor ATF4. Overexpression of mutated ATF4 that lacks the transcriptional activity did not alter EPO transcriptional regulation. Transcriptional activity of EPO 3' enhancer, which is mainly regulated by HIF, was abolished by both ER stressors and ATF4 overexpression, while nuclear HIF accumulation or expression of other HIF target genes was not suppressed by ER stress. Chromatin immunoprecipitation analysis identified a novel ATF4 binding site (TGACCTCT) within the EPO 3' enhancer region, suggesting a distinct role for ATF4 in UPR-dependent suppression of the enhancer. Induction of ER stress in rat liver and kidney by tunicamycin decreased the hepatic and renal mRNA and plasma level of EPO. Collectively, ER stress selectively impairs the transcriptional activity of EPO but not of other HIF target genes. This effect is mediated by suppression of EPO 3' enhancer activity via ATF4 without any direct effect on HIF, indicating that UPR contributes to oxygen-sensing regulation of EPO.

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    • "Aldolase (Ald) transcription start site resulted in clear Atf4-dependent up-regulation of GFP during ER stress (Figure 5A), but mutation of the 2 CREs within this reporter had no effect on its regulation (Figure 5B). These results open up the possibility that Atf4 also regulates glycolytic genes through alternative binding sites (Fawcett et al. 1999; Gombart et al. 2007; Gjymishka et al. 2008; Kode et al. 2012; Chiang et al. 2013; Han et al. 2013) or via both direct and indirect mechanisms. "
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    ABSTRACT: Endoplasmic reticulum (ER) stress results from an imbalance between the load of proteins entering the secretory pathway and the ability of the ER to fold and process them. The response to ER stress is mediated by a collection of signaling pathways termed the unfolded protein response (UPR), which plays important roles in development and disease. Here we show that in Drosophila melanogaster S2 cells, ER stress induces a coordinated change in the expression of genes involved in carbon metabolism. Genes encoding enzymes that carry out glycolysis were up-regulated, whereas genes encoding proteins in the TCA cycle and respiratory chain complexes were down-regulated. The UPR transcription factor Atf4 was necessary for the up-regulation of glycolytic enzymes and Lactate dehydrogenase (Ldh). Furthermore, Atf4 binding motifs in promoters for these genes could partially account for their regulation during ER stress. Finally, flies up-regulated Ldh and produced more lactate when subjected to ER stress. Together these results suggest that Atf4 mediates a shift from a metabolism based on oxidative phosphorylation to one more heavily reliant on glycolysis, reminiscent of aerobic glycolysis or the Warburg effect observed in cancer and other proliferative cells. Copyright © 2015 Author et al.
    G3-Genes Genomes Genetics 02/2015; 5(4). DOI:10.1534/g3.115.017269 · 3.20 Impact Factor
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    • "sion of transcriptio n factors, pCAX-F-hXBP1 (sp) [21], pCAX-hATF6(1-373)-F [30], and pCAX-F-mATF 4 [31] were used. In the reporter assays, pGL4.10 (Promega) was used as a negative control without promoter. "
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    ABSTRACT: IRE1 α is an ER-located transmembrane RNase whose activation leads to the production of the transcriptional factor, XBP1. Recently, many studies report that IRE1 α-XBP1 pathway has novel and significant roles in placenta. However, its molecular details have been still unknown. To address this point, we have focused on the molecular linkage between IRE1 α -XBP1 pathway and Cts7 and Cts8, which are essential cathepsins for placenta formation. In cellular model, this pathway positively contributed to their expression at transcriptional level. In addition, the disruption of IRE1 α or XBP1 in animal model significantly attenuated their transcripts in placenta. These results indicated that IRE1 α -XBP1 pathway function as a specific program supporting the placenta formation by ensuring the moderate expression of specific subset of placental cathepsins.
    Biochemical and Biophysical Research Communications 03/2013; 433(4). DOI:10.1016/j.bbrc.2013.02.119 · 2.30 Impact Factor
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    ABSTRACT: Experimental evidence suggests an immense variety of processes associated with and aimed at producing reactive oxygen and/or nitrogen species. Clinical studies implicate an enormous range of pathologies associated with reactive oxygen/nitrogen species metabolism deregulation, particularly oxidative stress. Recent advances in biochemistry, proteomics and molecular biology/biophysics of cells suggest oxidative stress to be an endpoint of complex dysregulation events of conjugated pathways consolidated under the term, proposed here, "oxidative status". The oxidative status concept, in order to allow for novel diagnostic and therapeutic approaches, requires elaboration of a new logic system comprehending all the features, versatility and complexity of cellular pro- and antioxidative components of different nature. We have developed a curated and regularly updated interactive interactome map of human cellular-level oxidative status allowing for systematization of the related most up-to-date experimental data. A total of more than 600 papers were selected for the initial creation of the map. The map comprises more than 300 individual factors with respective interactions, all subdivided hierarchically for logical analysis purposes. The pilot application of the interactome map suggested several points for further development of oxidative status-based technologies. Current OSIM release is available at
    Molecular BioSystems 05/2013; DOI:10.1039/c3mb70096h · 3.21 Impact Factor
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