• Title/Summary/Keyword: stress gene

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Protopanaxatriol Ginsenoside Rh1 Upregulates Phase II Antioxidant Enzyme Gene Expression in Rat Primary Astrocytes: Involvement of MAP Kinases and Nrf2/ARE Signaling

  • Jung, Ji-Sun;Lee, Sang-Yoon;Kim, Dong-Hyun;Kim, Hee-Sun
    • Biomolecules & Therapeutics
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    • v.24 no.1
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    • pp.33-39
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    • 2016
  • Oxidative stress activates several intracellular signaling cascades that may have deleterious effects on neuronal cell survival. Thus, controlling oxidative stress has been suggested as an important strategy for prevention and/or treatment of neurodegenerative diseases. In this study, we found that ginsenoside Rh1 inhibited hydrogen peroxide-induced reactive oxygen species generation and subsequent cell death in rat primary astrocytes. Rh1 increased the expression of phase II antioxidant enzymes, such as heme oxygenase-1 (HO-1), NAD(P)H:quinone oxidoreductase 1, superoxide dismutase-2, and catalase, that are under the control of Nrf2/ARE signaling pathways. Further mechanistic studies showed that Rh1 increased the nuclear translocation and DNA binding of Nrf2 and c-Jun to the antioxidant response element (ARE), and increased the ARE-mediated transcription activities in rat primary astrocytes. Analysis of signaling pathways revealed that MAP kinases are important in HO-1 expression, and act by modulating ARE-mediated transcriptional activity. Therefore, the upregulation of antioxidant enzymes by Rh1 may provide preventive therapeutic potential for various neurodegenerative diseases that are associated with oxidative stress.

Molecular cloning and characterization of a soybean GmMBY184 induced by abiotic stresses

  • Chung, Eun-Sook;Kim, Koung-Mee;Lee, Jai-Heon
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.175-181
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    • 2012
  • Drought and high salinity stresses often imposes adverse effects on crop yield. MYB transcription factors have been shown to be an important regulator in defense responses to these environmental stresses. In this study, we have cloned and characterized a soybean gene GmMYB184 (Glycine max MYB transcription factor 184). Deduced amino acid sequences of GmMYB184 show highest homology with that from Vitis vinifera legume plant (75%). Different expression patterns of GmMYB184 mRNA were observed subjected to drought, cold, high salinity stress and abscisic acid treatment, suggesting its role in the signaling events in the osmotic stress-related defense response. Subcellular localization studies demonstrated that the GFP-GmMYB184 fusion protein was localized in the nucleus. Using the yeast assay system, the C-terminal region of GmMYB184 was found to be essential for the transactivation activity. These results indicate that the GmMYB184 may play a role in abiotic stress tolerance in plant.

STATE-OF-THE-ART TECHNOLOGY USING GENETICALLY-ENGINEERED BIOLUMINESCENT BACTERIA AS ENVIRONMENTAL BIOSENSORS

  • Gu, Man-Bock
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.94-99
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    • 2000
  • Bioluminescence is being used as a prevailing reporter of gene expression in microorganisms and mammalian cells. Bacterial bioluminescence draws special attention from environmental biotechnologists since it has many advantageous characteristics, such as no requirement of extra substractes, highly sensitive, and on-line measurability. Using bacterial bioluminescence as a reporter of toxicity has replaced the classical toxicity monitoring technology of using fish or daphnia with a cutting-edge technology. Fusion of bacterial stress promoters, which control the transcription of stress genes corresponding to heat-shock, DNA-, or oxidative-damaging stress, to the bacterial lux operon has resulted in the development of novel toxicity biosensors with a short measurement time, enhanced sensitivity, and ease and convenient usage. Therefore, these recombinant bioluminescent bacteria are expected to induce bacterial bioluminescence when the cells are exposed to stressful conditions, including toxic chemicals. We have used these recombinant bioluminescent bacteria in order to develop toxicity biosensors in a continuous, portable, or in-situ measurement from for air, water, and soil environments. All the data obtained from these toxicity biosensors for these environments were found to be repeatable and reproducible, and the minimum detection level of toxicity was found to be ppb (part per billion) levels for specific chemicals.

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Expression of Arabidopsis Phytochelatin Synthase 2 Is Too Low to Complement an AtPCS1-defective Cad1-3 Mutant

  • Lee, Sangman;Kang, Beom Sik
    • Molecules and Cells
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    • v.19 no.1
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    • pp.81-87
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    • 2005
  • Phytochelatins play an important role in heavy metal detoxification in plants as well as in other organisms. The Arabidopsis thaliana mutant cad1-3 does not produce detectable levels of phytochelatins in response to cadmium stress. The hypersensitivity of cad1-3 to cadmium stress is attributed to a mutation in the phytochelatin synthase 1 (AtPCS1) gene. However, A. thaliana also contains a functional phytochelatin synthase 2 (AtPCS2). In this study, we investigated why the cad1-3 mutant is hypersensitive to cadmium stress despite the presence of AtPCS2. Northern and Western blot analyses showed that expression of AtPCS2 is weak compared to AtPCS1 in both roots and shoots of transgenic Arabidopsis. The lower level of AtPCS2 expression was confirmed by RT-PCR analysis of wild type Arabidopsis. Moreover, no tissue-specific expression of AtPCS2 was observed. Even when AtPCS2 was under the control of the AtPCS1 promoter or of the cauliflower mosaic virus 35S promoter (CaMV 35S) it was not capable of fully complementing the cad1-3 mutant for cadmium resistance.

A Lipid-derived Endogenous Inducer of COX-2: a Bridge Between Inflammation and Oxidative Stress

  • Uchida, Koji
    • Molecules and Cells
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    • v.25 no.3
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    • pp.347-351
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    • 2008
  • Several lines of evidence indicate that the oxidative modification of protein and the subsequent accumulation of the modified proteins have been found in cells during aging, oxidative stress, and in various pathological states including premature diseases, muscular dystrophy, rheumatoid arthritis, and atherosclerosis. The important agents that give rise to the modification of a protein may be represented by reactive aldehydic intermediates, such as ketoaldehydes, 2-alkenals and 4-hydroxy-2-alkenals. These reactive aldehydes are considered important mediators of cell damage due to their ability to covalently modify biomolecules, which can disrupt important cellular functions and can cause mutations. Furthermore, the adduction of aldehydes to apolipoprotein B in low-density lipoproteins (LDL) has been strongly implicated in the mechanism by which LDL is converted to an atherogenic form that is taken up by macrophages, leading to the formation of foam cells. During the search for an endogenous inducer of cyclooxygenase-2 (COX-2), an inducible isoform responsible for high levels of prostaglandin production during inflammation and immune responses, 4-hydroxy-2-noennal (HNE), one of the most representative lipid peroxidation product, has been identified as the potential inducer of COX-2. In addition, the following study on the molecular mechanism of the COX-2 induction by HNE has unequivocally established that a serum component, which is eventually identified to be denatured LDL, is essential for COX-2 induction. Here I review current understanding of the mechanisms by which HNE in cooperation with the serum component activates gene expression of COX-2.

Repressed Quorum Sensing by Overexpressing LsrR Hampers Salmonella Evasion from Oxidative Killing Within Macrophages

  • Choi, Jeong-Joon;Park, Joo-Won;Ryu, Sang-Ryeol
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1624-1629
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    • 2010
  • Bacterial cell-to-cell communication, termed quorum sensing (QS), leads to coordinated group behavior in a cell-density-dependent fashion and controls a variety of physiological processes including virulence gene expression. The repressor of the lsr operon, LsrR, is the only known regulator of LuxS/AI-2-mediated QS in Salmonella. Although lack of lsrR did not result in noticeable differences in Salmonella survival, the down-regulation of QS as a result of lsrR overexpression decreased Salmonella survival within macrophages. We found that impaired growth of Salmonella overexpressing lsrR within macrophages was due largely to its hypersensitivity to NADPH-dependent oxidative stress. This, in turn, was a result of decreased expression of genes involved in the oxidative stress response, such as sodA, sodCI, and sodCII, when lsrR was overexpressed. These results suggest that down-regulation of QS by excess LsrR can lower Salmonella virulence by hampering Salmonella evasion from oxidative killing within macrophages.

Molecular characterization of BrRZFPs genes encoding C3HC4 type RING zinc finger protein under abiotic stress from Chinese cabbage (Brassica rapa L.)

  • Jung, Yu Jin;Lee, Kye Dong;Cho, Yong Gu;Nou, Ill Sup;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.40 no.2
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    • pp.102-110
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    • 2013
  • The novel BrRZFPs genes encoding C3HC4-type RING zinc finger protein were identified from FOX (full length cDNA over-expressing) library of Brassica rapa. Ten full-length cDNAs obtained from the library encode zinc-finger protein containing 346 amino acids, designated BrRZFPs. These genes were classified into four groups by phylogenic analysis showing conserved protein sequences at both termini. The tissue distribution of BrRZFPs transcription was examined by qRT-PCR revealing ubiquitous expression pattern. However, each gene was strongly expressed in the specific tissue. Transcriptional analysis showed that those acquired 10 genes were inducible under abiotic stresses. Likewise, the transcript of BrRZFP3 was strongly induced (~12-folds) by exogenous abscisic acid, whereas the transcripts of BrRZFP1, BrRZFP2 and BrRZFP3 were (> 9-folds) induced by cold. We suggest that these BrRZFPs that function as signal or response to abiotic stress are useful for crop improvement.

Antioxidant Activity of Ethyl acetate Fraction of Oat in Caenorhabditis elegans (귀리 Ethyl acetate 분획물의 예쁜 꼬마선충 내의 항산화 효과)

  • Kwon, Kang Mu;Kim, Jun Hyeong;Yang, Jae Heon;Ki, Byeolhui;Hwang, In Hyun;Kim, Dae Keun
    • Korean Journal of Pharmacognosy
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    • v.52 no.4
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    • pp.251-256
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    • 2021
  • Oat, the seeds of Avena sativa L. (Gramineae), is an important dietary staple for people in many countries. Previous studies reported that A. sativa had various pharmacological effects such as anti-inflammatory, antitumor, neurotonic, and antispasmodic activities. In this study, Caenorhabditis elegans model system was used to investigate the antioxidant activity of methanol extract of oat. The ethyl acetate soluble fraction of the oat methanol extract showed the best DPPH radical scavenging activity. The ethyl acetate fraction was measured for the activity of superoxide dismutase (SOD), catalase, and oxidative stress tolerance by using C. elegans along with reactive oxygen species (ROS) level. In addition, to confirm that the regulation of the stress response gene is responsible for the increased stress tolerance of C. elegans treated by the ethyl acetate fraction, SOD-3 expression was measured using GFP-expressing transgenic worm. As a result, the ethyl acetate fraction increased SOD and catalase activities, and decreased ROS accumulation in a dose-dependent manner. In addition, the ethyl acetate fraction-treated CF1553 worm showed higher SOD-3::GFP intensity compared to the control.

Antioxidant Activities of n-Butanol Fraction of Pinus densiflora Siebold & Zucc. Needle in Caenorhabditis elegans. (솔잎 부탄올 분획의 예쁜꼬마선충 내의 항산화 효과)

  • Kim, Jun Hyeong;Kim, Na Yeong;Kwon, Kang Mu;Hwang, In Hyun;Kim, Dae Keun
    • Korean Journal of Pharmacognosy
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    • v.52 no.2
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    • pp.105-111
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    • 2021
  • To investigate the antioxidant activities of Pinus densiflora needle (Pinaceae), which is widely used as a traditional medicine in Korea, Caenorhabditis elegans model system was used. The n-butanol fraction of P. densiflora needle methanol extract showed the most potent DPPH radical scavenging and superoxide quenching activities in a dose-dependent manner. The n-butanol fraction was measured for the activities of superoxide dismutase (SOD), catalase, and oxidative stress tolerance by using C. elegans with reactive oxygen species (ROS) level. In addition, to see that the regulation of the stress response gene is responsible for the increased stress tolerance of C. elegans treated by the butanol extract, SOD-3 expression was measured using a transgenic strain. As a result, the P. densiflora needle n-butanol fraction increased SOD and catalase activities, and decreased ROS accumulation, dose-dependently. Furthermore, the n-butanol fraction-treated CF1553 worm showed higher SOD-3::GFP intensity than the control.

Antioxidant Activity of Helianthus tuberosus L. Flower in Caenorhabditis elegans (돼지감자꽃의 예쁜꼬마선충 내의 항산화 효과)

  • Kim, Jun Hyeong;Lee, Ye Bin;Han, Seon Yeo;Kim, Su Jin;Hwang, In Hyun;Kim, Dae Keun
    • Korean Journal of Pharmacognosy
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    • v.50 no.2
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    • pp.96-101
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    • 2019
  • Methanol extract of Helianthus tuberosus L. (Compositae) flower was investigated to research the anti-oxidative activity by using a Caenorhabditis elegans model system. Ethyl acetate soluble fraction of the methanol extract showed the most potent DPPH radical scavenging activity. The ethyl acetate fraction was measured on its activities of superoxide dismutase (SOD), catalase, and oxidative stress tolerance with reactive oxygen species (ROS) level in C. elegans. Furthermore, in order to verify if regulation of stress-response gene is responsible for the increased stress tolerance of C. elegans which treated by the ethyl acetate fraction, we checked SOD-3 expression using a transgenic strain. Consequently, the ethyl acetate fraction of H. tuberosus flower increased the catalase and SOD activities in a dose-dependent manner in C. elegans, reduced ROS accumulation dose-dependently. Besides, the ethyl acetate fraction-treated CF1553 worms showed higher SOD-3::GFP intensity than the control group.