Identification of an additional hypoxia responsive element in the glyceraldehyde-3-phosphate dehydrogenase gene promoter

Lu, Shan; Gu, Xiang; Hoestje, Sara M; Epner, Daniel E.

doi:10.1016/s0167-4781(01)00359-1

Cited by 78 publications

(50 citation statements)

References 20 publications

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“…Focusing on those genes that regulate the energetic state of nucleus pulposus cells, we assessed expression of the glucose transporters GLUT-1 and GLUT-3. In the same study, we also evaluated another HIF-1 target, GAPDH, the enzyme that catalyzes the conversion of glyceraldehyde 3-phosphate into 1,3-bisphosphoglycerate (13). At 2 and 20% O 2 , GLUT-1, GLUT-3, and GAPDH mRNA expression levels were comparable and remained constant over time.…”

Section: Discussionmentioning

confidence: 99%

“…Luciferase reporter plasmids were provided by Dr. Sherin Devaskar, University of California, Los Angels (Glut-3 promoter) (19), Dr. Oleksandr Minchenko, Palladin Institute of Biochemistry, Ukraine [6-phosphofructo-2-kinase/fructose-2,6-biphosphatase (PFKFB) promoter and HIF-1␣ 3Ј-untranslated region (UTR)] (14), and Dr. Shan Lu, University of Cincinnati (GAPDH promoter) (13). pGlut-1 wild type (Wt) and pGlut-1 mutant (Mt) were provided by Dr. Amit Maity, University of Pennsylvania (4); pGlut-1 (Wt) contains an active HRE, whereas the 4-bp mutation in pGlut-1 (Mt) disrupts HIF-1 binding.…”

Section: Examination Of Hif-1␣ and Glut-1 Expression By Immunofluoresmentioning

confidence: 99%

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Normoxic stabilization of HIF-1α drives glycolytic metabolism and regulates aggrecan gene expression in nucleus pulposus cells of the rat intervertebral disk

Agrawal¹,

Guttapalli²,

Narayan³

et al. 2007

American Journal of Physiology-Cell Physiology

154

180

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Agrawal A, Guttapalli A, Narayan S, Albert TJ, Shapiro IM, Risbud MV. Normoxic stabilization of HIF-1␣ drives glycolytic metabolism and regulates aggrecan gene expression in nucleus pulposus cells of the rat intervertebral disk. Am J Physiol Cell Physiol 293: C621-C631, 2007. First published April 18, 2007; doi:10.1152/ajpcell.00538.2006.-The nucleus pulposus is an aggrecan-rich, avascular tissue that permits the intervertebral disk to resist compressive loads. In the disk, nucleus pulposus cells express hypoxia-inducible factor (HIF)-1␣, a transcription factor that responds to oxygen tension and regulates glycolysis. The goal of the present study was to examine the importance of HIF-1␣ in rat nucleus pulposus cells and to probe the function of this transcription factor in terms of regulating aggrecan gene expression. We found that HIF-1␣ protein levels and mRNA stability were similar at 20 and 2% O2; there was a small, but significant increase in HIF-1␣ transactivation domain activity in hypoxia. With respect to HIF-1␣ target genes GAPDH, GLUT-1, and GLUT-3, mRNA and protein levels were independent of the oxygen tension. Other than a modest increase in 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase reporter activity, the oxemic state did not change GAPDH, GLUT-1, and GLUT-3 promoter activities. Treatment of cells with 2-deoxyglucose (2-DG), a glycolytic inhibitor, resulted in a significant suppression in ATP synthesis in normoxia, whereas treatment with mitochondrial inhibitors did not affect ATP production and cell viability. However, measurement of the rate of fatty acid oxidation indicated that these cells contained functioning mitochondria. Finally, we showed that when HIF-1␣ was suppressed, irrespective of the oxemic state, there was a partial loss of aggrecan expression and promoter activity. Moreover, when cells were treated with 2-DG, there was inhibition in aggrecan promoter activity. Results of this study indicate that oxygen-independent stabilization of HIF-1␣ in nucleus pulposus cells is a metabolic adaptation that drives glycolysis and aggrecan expression. hypoxia; hypoxia-inducible factor-1␣ THE LOCAL OXYGEN TENSION promotes cellular differentiation and regulates tissue energy metabolism. When the oxygen tension is low, there is almost complete reliance on glycolysis to generate ATP and reducing equivalents (12,29,30). In specialized tissues such as the intervertebral disk, the mechanism by which cells conserve energy has received little attention. Nonetheless, it is known that in the intervertebral disk, the nucleus pulposus has no direct vascular supply (8,24,33), and ATP is probably generated by anaerobic glycolysis (1, 10). The dependence on glycolysis reflects the low oxygen tension in the disk, which has been reported to be low as low as 2%, and the observation that modulation in the oxygen supply can cause vertebral defects (15). Regulation of glycolysis is mediated by hypoxia-inducible factor (HIF)-1␣, a transcription factor that responds to the local oxygen tension, and has been show...

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Section: Discussionmentioning

confidence: 99%

Section: Examination Of Hif-1␣ and Glut-1 Expression By Immunofluoresmentioning

confidence: 99%

Normoxic stabilization of HIF-1α drives glycolytic metabolism and regulates aggrecan gene expression in nucleus pulposus cells of the rat intervertebral disk

Agrawal¹,

Guttapalli²,

Narayan³

et al. 2007

American Journal of Physiology-Cell Physiology

154

180

View full text Add to dashboard Cite

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“…Because the Gapdh promoter region contains hypoxia-responsive elements (22), exposure of mice we studied to a hypoxic environment might alter the expression of Gapdh. Therefore, we measured pulmonary Gapdh expression (relative to actin) in the lungs of Cyp2j 2/2 and Cyp2j 1/1 male mice exposed to hypoxia and normoxia for 6 weeks.…”

Section: Rv Hypertrophymentioning

confidence: 99%

Pulmonary Hypertension after Prolonged Hypoxic Exposure in Mice with a Congenital Deficiency of Cyp2j

Beloiartsev

Rodrigues‐Machado

Zhou

et al. 2015

Am J Respir Cell Mol Biol

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Cytochrome P450 epoxygenase-derived epoxyeicosatrienoic acids contribute to the regulation of pulmonary vascular tone and hypoxic pulmonary vasoconstriction. We investigated whether the attenuated acute vasoconstrictor response to hypoxic exposure of Cyp2j 2/2 mice would protect these mice against the pulmonary vascular remodeling and hypertension associated with prolonged exposure to hypoxia. Cyp2j 2/2 and Cyp2j 1/1 male and female mice continuously breathed an inspired oxygen fraction of 0.21 (normoxia) or 0.10 (hypoxia) in a normobaric chamber for 6 weeks. We assessed hemoglobin (Hb) concentrations, right ventricular (RV) systolic pressure (RVSP), and transthoracic echocardiographic parameters (pulmonary acceleration time [PAT] and RV wall thickness). Pulmonary Cyp2c29, Cyp2c38, and sEH mRNA levels were measured in Cyp2j 2/2 and Cyp2j 1/1 male mice. At baseline, Cyp2j 2/2 and Cyp2j 1/1 mice had similar Hb levels and RVSP while breathing air. After 6 weeks of hypoxia, circulating Hb concentrations increased but did not differ between Cyp2j 2/2 and Cyp2j 1/1 mice. Chronic hypoxia increased RVSP in Cyp2j 2/2 and Cyp2j 1/1 mice of either gender. Exposure to chronic hypoxia decreased PAT and increased RV wall thickness in both genotypes and genders to a similar extent. Prolonged exposure to hypoxia produced similar levels of RV hypertrophy in both genotypes of either gender. Pulmonary Cyp2c29, Cyp2c38, and sEH mRNA levels did not differ between Cyp2j 2/2 and Cyp2j 1/1 male mice after breathing at normoxia or hypoxia for 6 weeks. These results suggest that murine Cyp2j deficiency does not attenuate the development of murine pulmonary vascular remodeling and hypertension associated with prolonged exposure to hypoxia in mice of both genders.Keywords: hypoxia; cytochrome P450 epoxygenases; pulmonary vascular remodeling; right ventricular hypertrophy Pulmonary hypertension (PH) due to increased vascular tone in lung vessels is important in many acute and chronic lung diseases, such as pulmonary embolism and idiopathic pulmonary arterial hypertension. Although progress has been made with therapeutic approaches to reduce vascular tone, all of the pathways influencing lung vascular tone have not been fully explored.

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“…An experimental model of BOO was established as described in Materials and Methods, and after 24 h, expression of endogenous indicators for individual stress was evaluated by northern blot analysis. The indicators used were COX-2 for mechanical stress, 17 GAPDH and VEGF for hypoxic stress [18][19][20] and GRP78 and CHOP for ER stress. 7,21 Northern blot analysis showed that, compared with untreated and sham-operated rats, rats subjected to BOO exhibited induction of COX-2 (Figure 1a), GAPDH and VEGF (Figure 1b) in the bladders.…”

Section: Induction Of Mechanical Stress Hypoxic Stress and Er Stressmentioning

confidence: 99%

Involvement of hypoxia-triggered endoplasmic reticulum stress in outlet obstruction-induced apoptosis in the urinary bladder

Sawada

Yao

Hiramatsu

et al. 2008

Laboratory Investigation

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In bladder outlet obstruction (BOO), mechanical stress and ischemia/hypoxia are implicated in structural and functional alterations of the urinary bladder. Because mechanical stress and hypoxia may trigger endoplasmic reticulum (ER) stress, we examined involvement of ER stress in the damage of the bladder caused by BOO. An experimental model of BOO was established in rats by complete ligature of the urethra for 24 h, and bladders were subjected to northern blot analysis and assessment of apoptosis. Isolated urinary bladders and bladder-derived smooth muscle cells (BSMCs) were also exposed to mechanical strain and hypoxia and used for analyses. To examine involvement of ER stress in the damage of the bladder, the effects of a chemical chaperone 4-phenylbutyrate (4-PBA) were evaluated in vitro and in vivo. Outlet obstruction for 24 h induced expression of ER stress markers, GRP78 and CCAAT/enhancer-binding protein-homologous protein (CHOP), in the bladder. It was associated with induction of markers for mechanical stress (cyclooxygenases 2) and hypoxia (vascular endothelial growth factor and glyceraldehyde-3-phosphate dehydrogenase). When isolated bladders and BSMCs were subjected to mechanical strain, induction of GRP78 and CHOP was not observed. In contrast, when BSMCs were exposed to hypoxic stress caused by CoCl 2 or thenoyltrifluoroacetone (TTFA), substantial upregulation of GRP78 and CHOP was observed, suggesting involvement of hypoxia in the induction of ER stress. In the bladder subjected to BOO, the number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive cells increased in the epithelial cells and BSMCs. Similarly, treatment with TTFA or CoCl 2 induced apoptosis of BSMCs, and 4-PBA significantly attenuated ER stress and apoptosis triggered by these agents. Furthermore, in vivo administration with 4-PBA significantly reduced apoptosis in the bladder subjected to BOO. These results suggested that outlet obstruction caused ER stress via hypoxic stress in the bladder and that hypoxia-triggered ER stress may be involved in the induction of apoptosis in BOO.

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Identification of an additional hypoxia responsive element in the glyceraldehyde-3-phosphate dehydrogenase gene promoter

Cited by 78 publications

References 20 publications

Normoxic stabilization of HIF-1α drives glycolytic metabolism and regulates aggrecan gene expression in nucleus pulposus cells of the rat intervertebral disk

Normoxic stabilization of HIF-1α drives glycolytic metabolism and regulates aggrecan gene expression in nucleus pulposus cells of the rat intervertebral disk

Pulmonary Hypertension after Prolonged Hypoxic Exposure in Mice with a Congenital Deficiency of Cyp2j

Involvement of hypoxia-triggered endoplasmic reticulum stress in outlet obstruction-induced apoptosis in the urinary bladder

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