• Title/Summary/Keyword: ${\alpha}$-ketoglutarate

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HIF-1-Dependent Induction of Jumonji Domain-Containing Protein (JMJD) 3 under Hypoxic Conditions

  • Lee, Ho-Youl;Choi, Kang;Oh, Hookeun;Park, Young-Kwon;Park, Hyunsung
    • Molecules and Cells
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    • v.37 no.1
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    • pp.43-50
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    • 2014
  • Jumonji domain-containing proteins (JMJD) catalyze the oxidative demethylation of a methylated lysine residue of histones by using $O_2$, ${\alpha}$-ketoglutarate, vitamin C, and Fe(II). Several JMJDs are induced by hypoxic stress to compensate their presumed reduction in catalytic activity under hypoxia. In this study, we showed that an H3K27me3 specific histone demethylase, JMJD3 was induced by hypoxia-inducible factor (HIF)-$1{\alpha}/{\beta}$ under hypoxia and that treatment with Clioquinol, a HIF-$1{\alpha}$ activator, increased JMJD3 expression even under normoxia. Chromatin immunoprecipitation (ChIP) analyses showed that both HIF-$1{\alpha}$ and its dimerization partner HIF-$1{\beta}$/Arnt occupied the first intron region of the mouse JMJD3 gene, whereas the HIF-$1{\alpha}/{\beta}$ heterodimer bound to the upstream region of the human JMJD3, indicating that human and mouse JMJD3 have hypoxia-responsive regulatory regions in different locations. This study shows that both mouse and human JMJD3 are induced by HIF-1.

Hypoxia suffocates histone demethylases to change gene expression: a metabolic control of histone methylation

  • Park, Hyunsung
    • BMB Reports
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    • v.50 no.11
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    • pp.537-538
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    • 2017
  • Hypoxia affects various physiological and pathophyological processes. Hypoxia changes the expression of hypoxia-responsive genes through two main pathways. First, hypoxia activates transcription factors (TF) such as Hypoxia-inducible Factor (HIF). Second, hypoxia decreases the activity of Jumonji C domain-containing histone demethylases (JMJDs) that require $O_2$ and ${\alpha}$-Ketoglutarate (${\alpha}$-KG) as substrates. The JMJDs affect gene expression through their regulation of active or repressive histone methylations. Profiling of H3K4me3, H3K9me3, and H3K27me3 under both normoxia and hypoxia identified 75 TFs whose binding motifs were significantly enriched in the methylated regions of the genes. TFs showing similar binding strengths to their target genes might be under the 'metabolic control' which changes histone methylation and gene expression by instant changing catalytic activities of resident histone demethylases.

Effect of Dietary Protein and Taurine on Cysteine Catabolism in Cat Liver (식이내의 단백질과 타우린 함량이 Cysteine 대사에 미치는 영향)

  • 박태선
    • Journal of Nutrition and Health
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    • v.29 no.7
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    • pp.729-737
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    • 1996
  • Activieties of hepatic cysteine desulfhydration was assessed in cats fed one of the following diets for 5 weeks : 20% protein, 0% taurine diet(LPOT) ; 20% protein, 0.15% taurine diet (LPNT) ; 60% protein, 0% taurine diet(HPOT) ; and 60% protein, 0.15% taurine diet(HPNT). Cats fed LPOT and HPOT had been maintained on a taurine-free diet for 6 weeks prior to the experiment in order to deplete body taurine. Activities of cysteine desulfhydration were determined by measuring the production of H235S from 35S-cysteine in the presence and absence of $\alpha$-ketoglutarate ($\alpha$-KG) in the incubation medium. The direct pathway via cysteine desulfhydrase appears to account for the major route of cysteine desulfhydration in the cat liver since the values obtained in the absence of $\alpha$-KG were between 81 and 88% of those obtained in the presence of $\alpha$-KG. Mean$\pm$SEM of the hepatic total desulfhydration activities(umol H2S.min-1.kg body wt-1)in cats fed LPOT, LPNT, HPOT and HPNT were 117$\pm$6, 135$\pm$10, 137$\pm$10, and 190$\pm$9, respectively. The capacity of hepatic cysteine desulfhydration (UA/kg body wt) was positively cerrelated not only with the dietary concentration of taurine but also with the concentration of protein.

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Precursors for the Ethylene Evolution of Pseudornonas syringae pv. Phaseolicola (Pseudomonas syringae pv. Phaseolicola에 의한 Ethylene 생성에서의 전구물질)

  • Bae, Moo;Kweon, Hea-Young
    • Microbiology and Biotechnology Letters
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    • v.19 no.1
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    • pp.14-20
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    • 1991
  • - The purpose of this work is to investigate the effects of various substrates on biosynthesis of ethylene by the Kudzu strain of Pseudomonas syn'ngae pv. Phaseolicola causing halo blight. In the intact cell of P. sym'ngue, optimal condition for ethylene production was achieved at p1-I 7.5 and $30^{\circ}C$ for 9 to 10 hours of culture. Ethylene was most effectively produced from amino acids such as Asn, Gln, Asp ans Glu, compared to those of various kinds of sugars. While ethylene production from $\alpha$-ketoglutarate ($\alpha$-KG) was gradually increased throughout 51 hours incubation period tested. Ethylene production derived from citrate, $\alpha$-KG and oxalacetate as well as a few amino acids was further enhanced by the addition of histidine or arginine. In cell-free ethylene-forming system, ethylene was most effectively produced from $\alpha$-KG, compared to those from citrate, oxalacetate, Glu, Arg, or Asp, at 0.5 mM among the range from 0.25 mM to 5 mM. Anlinooxyacetate, an inhibitor of a pyridoxal phosphate-linked enzyme, completely inhibited ethylene evolution derived from Glu but not affect that derived from $\alpha$-KG. The results obtained in this work suggest that $\alpha$-KG might be a direct precursor of ethylene production in this organism than any other substrates tested.

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Effect of Cadmium on Organic Acid Transport System in Renal Basolateral Membrane

  • Kim, Ghi-Chan;Kim, Kyoung-Ryong;Kim, Jee-Yeun;Park, Yang-Saeng
    • The Korean Journal of Physiology
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    • v.30 no.2
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    • pp.279-288
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    • 1996
  • Chronic exposure to cadmium impairs various renal tubular functions, including organic acid (anion) secretion. To investigate the mechanism of cadmium-induced alterations in the organic anion transport system, kinetics of p-aminohippurate (PAH) uptake was studied in renal cortical basolateral membrane vesicles (BLMV) isolated from cadmium-intoxicated rats (adult male Sprague-Dawley). Cadmium intoxication was induced by subcutaneous injections of $CdCl_{2}$ (2 mg Cd/kg per day) for 3 weeks. The renal plasma membrane vesicles were prepared by Percoll gradient centrifugation. The vesicular uptake of $^{14}C$-PAH was determined by rapid filtration technique using Millipore filter. Cadmium intoxication resulted in a marked attenuation of $Na^{+}$-dependent, ${\alpha}$-ketoglutarate (${\alpha}$KG)-driven PAH uptake with no changes in $Na^{+}$ and ${\alpha}$KG-independent transport component. Kinetic analysis indicated that Vmax, but not Km, of the $Na^{+}$-dependent, ${\alpha}$KG-driven component was reduced. A similar reduction of $Na^{+}$-dependent, ${\alpha}$KG-driven PAH uptake was observed in normal membrane vesicles directly exposed to inorganic cadmium in vitro, and this was accompanied by an inhibition of both $Na^{+}$-dependent ${\alpha}$KG uptake and ${\alpha}$KG-PAH exchange activity. These results indicate that during chronic exposure to cadmium, free cadmium ions liberated in the proximal tubular cytoplasm directly interact with the basolateral membrane and impair the active transport capacity for organic anions, most likely due to an inhibition of both $Na^{+}$-dicarboxylate cotransporter and dicarboxylate-organic anion antiporter activities.

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Characteristics of a New Obligate Methanol-Oxidizing Bacterium (새로운 절대 메탄올 산화세균의 분리 및 특성)

  • Kim, Si-Wook;Park, Yong-Ha
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.261-266
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    • 1993
  • A new methyltrophic bacterium which utilizes methanol as a sole source of carbon and energy was isolated from soil. It was Gram-negative, nonmotile, nonspore-forming rod, and strictly aerobic bacterium. Catalase and oxidase activities were present. Nitrate was reduced to nitrite. Vitamins and other growth factors were not required. Generation time was 1.6 hr under the optimal condition. The isolate assimilated methanol via the ribulose mono-phosphate pathway (Enter-Doudoroff varient) and did not have .alpha.-ketoglutarate dehydrogenase. It assimilated ammonia through glutamate dehydrogenase. The guanine plus cytosine content of the DNA was 61.0 mol%. The celular fatty acid composition was primarily straight-chain saturated $C^{16 : 0}$ acids (palmitic acids) and unsaturated $C_{16 :1}$ acid (palmitoleic acids), and the isolate also contained two unidentified $C_{17}$ branched fatty acids. The major ubiquinone was Q-8, and Q-6 and Q-7 were present as minor components. Phosphatidylethanolamine and phosphatidylglycerol were predominantly present, and diphosphatidyglycerol was also detected. Based on the physiological and biochemical properties, the isolate was assigned to a novel species of the genus Methylobacillus, Methylobacillus methanolovorus sp. nov.

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Effect of KGD1 Deletion on Cell Wall Biogenesis in Saccharomyces cerevisiae (Saccharomyces cerevisiae의 KGD1 유전자 결손이 세포벽 생합성에 미치는 영향)

  • Kim, Sung-Woo;Ahn, Ki-Woong;Park, Yun-Hee;Park, Hee-Moon
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.29-33
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    • 2010
  • KGD1 gene was cloned by functional complementation of defects in $\beta$-1,3-glucan synthase activity of the previously isolated Saccharomyces cerevisiae mutant LP0353, which displays a number of cell wall defects at restrictive temperature. We performed the gene disruption experiment to characterize the function of KGD1 gene, which encodes $\beta$-ketoglutarate dehydrogenase, in cell wall biosynthesis. The disruption of KGD1 showed the decreased growth rate, the increase of chitin synthases activity, alterations in cell wall composition, and increase of susceptibility to cell wall inhibitors such as Calcofluor white and Nikkomycin Z. These results suggested that KGD1 might be involved in cell wall biogenesis, especially the biosynthesis of $\beta$-1,6-glucan and chitin in S. cerevisaie.

Inhibitory Actions of Mycotoxins on Brain $\gamma$-Aminobutyrate Transaminase ($\gamma$-Aminobutyrate Transaminase에 대한 Mycotoxin의 저해작용)

  • Lee, Su-Jin;Lee, Kil-Soo;Choi, Soo-Young
    • Korean Journal of Microbiology
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    • v.31 no.3
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    • pp.224-229
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    • 1993
  • GABA transminase (4-aminobutyrate aminotransferase), which catalyzes the breakdown of the major inhibitory neurotransmitter, GABA, in mammalian brain, was inactivated by preincubation with the mycotoxin patulin. The time course of the reaction was significantly affected by the substrate .alpha.-ketoglutarate, which aforded complete protection against the loss of catalytic activity. The recovery from the inhibition of patulin by the addition of dithiothreitol (DTT) supports that patulin reacts with the sulfhydryl residue in the catalytic domain of the enzyme. The reconstitution of the reduced enzyme and apoenzyme with pyridoxal-5-P(PLP) was inhibited by another mycotoxin, penicilic acid. This mycotoxin may interact with lysyl residue of the enzyme. Therefore, it is postulated that the critical sulfhydryl and lysyl residues in the catalytic domain of the enzyme react with mycotoxin patulin and penicillic acid, respectively.

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Genetic regulation for the biosynthesis of glutamate family in Corynebacterium glutamicum (Corynebacterium glutamicum에서의 glutamate계 아미노산 생합성의 유전적 조절)

  • Kim In-Ju;Kyung Hee Min;Sae Bae Lee
    • Microbiology and Biotechnology Letters
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    • v.14 no.5
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    • pp.427-432
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    • 1986
  • The regulation of three ammonia assimilatory enzymes, GDH (glutamate dehydrogenase), GS (glutamine synthetase) and GOGAT (glutamate synthase), has been examined in C. glutamicum. Three kinds of arginine auxotrophs blocked in each step of arginine biosynthetic pathway from glutamate were selected as arg 5, arg 6, arg 8. Histidine and tryptophan auxotrophs were also selected because histidine and tryptophan repressed GS biosynthesis in E. coli. These strains were cultured on the media containing nitrogen-excess and limited conditions, to compare the specific activities of ${\alpha}$-ketoglutarate dehydrogenase(${\alpha}-KGDH$), GDH, GS, GOGAT from the cell-free extracts. These results showed that enzyme levels of ${\alpha}-KGDH$ and GDH from 3 kinds of arginine auxotrophs, histidine and tryptophan auxotrophs in nitrogen-excess condition and those of GS and GOGAT in nitrogen limited condition were increased compared with opposite condition. The tryptophan and histidine auxotrophs showed higher level of glutamate and glutamine than parental strains and other mutants. it is assumed that the higher levels of ${\alpha-KGDH}$ and GDH from mutants in nitrogen-excess condition promoted the accumulation of glutamate and glutamine in fermentation broth. The inhibition of GS activities by ADP suggested that GS is regulated by energy charge in C. glutamicum. The results with histidine, tryptophan, glycine, alanine, serine and GMP implied that a system of feedback inhibition were effective. The GDH, GS and GOGAT biosynthesis in culture broth was markedly repressed by the nature and kinds of available nitrogen sources such as tryptophan, proline, glycine, alanine, serine and tyrosine.

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Isolation and Characteristics of ${\varepsilon}$-Caprolactam Utilizing Bacteria (${\varepsilon}$-Caprolactam 이용성(利用性) 세균(細菌)의 분리(分離) 및 그 성질(性質))

  • Choi, Sun Taek;Rhee, In Koo
    • Current Research on Agriculture and Life Sciences
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    • v.3
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    • pp.21-27
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    • 1985
  • A bacterium which utilizes ${\varepsilon}$-caprolactam as a sole source of carbon and nitrogen was isolated from sludge of Shinchun river in Taegu and identified as Arthrobacter globiformis N-2-l. The growth medium for the optimum culture condition was composed of 0.4% ${\varepsilon}$-caprolactam, 0.02% $K_2HPO_4$, 0.05% $KH_2PO_4$, 0.02% $MgSO_4{\cdot}7H_2O$, 0.01% $FeCl_3{\cdot}6H_2O$ and 0.05% yeast extract. The optimum pH and temperature for growth were 7.0 and $30^{\circ}C$ respectively. The bacterial growth on the ${\varepsilon}$-caprolactam medium did not require any other organic nitrogen source such as yeast extract, although it was remarkably stimulated by the yeast extract. The bacteria utilized wide range of sugars and organic acids such as ${\alpha}$-ketoglutarate, adipate and P-hydroxybenzoate. The bacteria could use all kind of amino acids, ${\varepsilon}$-Caprolactam in the medium was consumed completely in the timecourse culture at $30^{\circ}C$ for 60 hr on the shaker by the bacteria. Decomposition product of ${\varepsilon}$-caprolactam by Arthrobacter globiformis N-2-1 was ${\varepsilon}$-aminocaproic acid.

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