• BMB Reports
• /
• v.39 no.3
• /
• pp.304-310
• /
• 2006

Transcription factor NF-E2-related factor 2 (Nrf2) regulates the induction of Phase II detoxifying enzymes and antioxidant enzymes in response to many cancer chemopreventive compounds. In this study, we investigated the role of receptor associated coactivator (RAC3) or steroid receptor coactivator-3 (SRC3) and other nuclear co-regulators including CBP/p300 (CREB-binding protein), CARM1 (Coactivator-associated arginine methyltransferase), PRMT1 (Protein arginine methyl-transferase 1), and p/CAF (p300/CBP-associated factor) in the transcriptional activation of a chimeric Gal4-Nrf2-Luciferase system containing the transactivation domain (TAD) of Nrf2 in HepG2 cells. The results indicated that RAC3 up-regulated the transactivation activity of Gal4-Nrf2-(1-370) in a dose-dependent manner. The enhancement of transactivation domain activity of Gal4-Nrf2-(1-370) by RAC3 was dampened in the presence of dominant negative mutants of RAC3. Next we studied the effects of other nuclear co-regulators including CBP/p300, CARM1, PRMT1 and p/CAF, and the results showed that they had different level of positive effects on this transactivation domain activity of Gal4-Nrf2-(1-370). But importantly, synergistic effects of these co-regulators in the presence of RAC3/SRC3 on the transactivation activity of Gal4-Nrf2-(1-370) were observed. In summary, our present study showed for the first time that the 160 RAC3/SRC3 is involved in the functional transactivation of TAD of Nrf2 and that the other nuclear co-regulators such as CBP/p300, CARM1, PRMT1 and p/CAF can also transcriptionally activate this TAD of Nrf2 and that they could further enhance the transactivation activity mediated by RAC3/SRC3.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.167-167
• /
• 2017

Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_ Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.105-105
• /
• 2017

Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

• Applied Biological Chemistry
• /
• v.38 no.4
• /
• pp.320-324
• /
• 1995

Solubilization of microsomal ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl transfearse(${\Delta}^{5}-3{\beta}$-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of ${\Delta}^{5}-3{\beta}$-OH-SAT and phosphatidylethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the ${\Delta}^{5}-3{\beta}$-OH SAT.

• The Korean Journal of Pesticide Science
• /
• v.10 no.1
• /
• pp.50-55
• /
• 2006

A field-resistant strain of the diamondback-moth(Plutella xylostella L.), collected from Chinese cabbage alpine farmland at Gangwon-do, Korea, was used for determination of the characteristics of resistance to chlorpyrifos using the activities of esterases and glutathione-S-transferase(GST), protein sequestration and AChE insenstivity. Although the activities of esterases extracted from resistant strain and susceptible strain were not significantly different, isozyme bands shown on the electrophoresis were different. GST activity from field resistant strain was 1.5-fold higher than that of susceptible. No differences were shown between resistant and susceptible ones in protein sequestration. The insensitivities of AChE to chlorpyrifos, however, extracted from susceptible strain was 460-fold higher than those of resistant. These results indicated that the insensitivity of AChE is the major factor for developing the resistance and activities of GST might be a minor factor.

• Journal of Life Science
• /
• v.18 no.7
• /
• pp.940-946
• /
• 2008

Neurotransmitter transporters, which remove neurotransmittesr from the synaptic cleft, are regulated by second messenger such as protein kinases and binding proteins. Neuronal ${\gamma}-aminobutyric$ acid transporters (GATs) are responsible for removing the inhibitory neurotransmitter ${\gamma}-aminobutyric$ acid (GABA) from the synaptic cleft. ${\gamma}-aminobutyric$ acid transporters 2 (GAT2/BGT1) is involved in regulating neurotransmitter recycling, but the mechanism how they are stabilized and regulated by the specific binding protein has not yet been elucidated. Here, we used the yeast two-hybrid system to identify the specific binding protein(s) that interacts with the C-terminal region of GAT2 and found a specific interaction with the mammalian LIN-7b (MALS-2). MALS-2 protein bound to the tail region of GAT2 but not to other GAT members in the yeast two-hybrid assay. The "T-X-L" motif at the C-terminal end of GAT2 is essential for interaction with MALS-2. In addition, this protein showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to GAT2 specifically co-immunoprecipitated MALS associated with GAT2 from mouse brain extracts. These results suggest that MALS may stabilize GAT2 in brain.

• Journal of Korean Society on Water Environment
• /
• v.32 no.5
• /
• pp.419-432
• /
• 2016

The aim of this study is to assess the influence of effluent discharge from a sewage treatment plant by evaluating oxidative stress and histopathological alterations in freshwater crucian carp Carassius auratus collected from the Eungcheon stream, located in Korea. Catalase activity in the gills, liver, and kidneys of C. auratus was collected from mixing zones; the downstream site was notably higher of fish than that of the upstream site. In addition, the activity of glutathione-S-transferase in the gills and liver was significantly higher in samples from the mixing zone than in those from the upstream site (p < 0.05). In addition, significantly elevated lipid peroxidation levels were observed in fish livers sampled from the mixing zone than in those from the upstream site (p < 0.05). Significant histopathological alternations were also observed in C. auratus, with the order of magnitude changes being liver > kidney > gills. These findings suggest that the liver is most affected by effluent discharge. The degree of tissue changes (DTC) indicate that the highest level occurred in samples from the mixing zone (30.98 ± 5.40) followed by those from the downstream site (19.28 ± 4.31) and was the lowest in samples from the upstream site (4.83 ± 2.67). These findings indicate that fish collected from the mixing zone are most affected by effluent discharge and both oxidative stress and histopathological indices are useful tools for monitoring contaminated rivers and streams.

• Journal of Life Science
• /
• v.19 no.8
• /
• pp.1055-1061
• /
• 2009

${\alpha}$-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors are widespread throughout the central nervous system and appear to serve as synaptic receptors for fast excitatory synaptic transmission mediated by glutamate. Their modulation is believed to affect learning and memory. To identify the interaction proteins for the AMPA receptor subunit glutamate receptor-interacting protein 1 (GRIPl), GRIP1 interactions with armadillo family protein p0071/plakophilin-4 were investigated. GRIP1 protein bound to the tail region of p0071/plakophilin-4 but not to other armadillo family protein members in a yeast two-hybrid assay. The "S-X-V" motif at the carboxyl (C)-terminal end of p0071/plakophilin-4 is essential for interaction with GRIP1. p0071/plakophilin-4 interacted with the Postsynaptic density-95/Discs large/Zona occludens-1 (PDZ) domains of GRIPI in the yeast two-hybrid assay, as is indicated also by Glutathione S-transferase (GST) pull-down, and co-immunoprecipitated with GRIP1 antibody in brain fraction. The findings of this study provide evidence that p0071/plakophilin-4 is an interactor of GRIP1.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.36 no.2
• /
• pp.159-166
• /
• 2016

This study was conducted to examine the effect of IgY supplementation on growth performance, blood metabolism, and disease occurrence in the weaned calves at 3 or 4 month of age. Average daily gain (ADG) and feed intakes were not affected by IgY supplementation, regardless of weaning months. White blood cell (WBC) count of the control group was higher in the final stage than in the initial stage (p<0.05), while that of the IgY supplemented group was lower in the final stage compared to the initial stage (p<0.05). regardless of treatments. Hematocrit (HCT) and red blood cell (RBC) counts were lower in the final stage than in the initial stage (p<0.05), but mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) counts were higher in the final stage than in the initial stage, regardless of treatments. In the control group, concentrations of albumin and globulin were higher in the final stage compared to the initial stage in the weaned calves at 3 month of age (p<0.05), and concentration of blood urea nitrogen (BUN) was increased in the IgY supplemented group. In the control group, concentrations of aspartate aminotransferase (AST) and gamma (${\gamma}$)-glutamyl transferase (GGT) were higher in the final stage compared to the initial stage (p<0.05). In contrast, concentrations of aspartate aminotransferase (AST) and gamma(${\gamma}$)-glutamyl transferase (GGT) were lower in the final stage compared to the initial stage for the IgY supplemented group. A diarrhea calf was only found in the control group. Manure score was higher in the control group compared to IgY supplemented group (p<0.05). Thus, these results indicates that IgY supplementation had positive effects on some blood metabolites, fecal condition score and diarrhea without negative effect on growth performance of the weaned Hanwoo calves.

• The Korean Journal of Mycology
• /
• v.30 no.2
• /
• pp.136-141
• /
• 2002

A hot water extract was prepared from the artificially grown Gastrodia elata to investigate its chemical composition and various in vitro biological activities as an effort to develop G. elata as health/functional food materials. The contents of crude protein, ash, fat, fiber, moisture and total sugar were 5.4, 2.6, 3.6, 3.3, 8.1 and 77% (w/w), respectively. The extract of G. elata had greater amount of potassium (1,150 mg/100 g) than phosphorus (300 mg/100 g). Dose-dependence against human carcinoma (Hep3B, MCF-7, A549 and AGS) were observed from 0.2 mg/ml to 1.0 mg/ml. Especially, the treatment of 1.0 mg/ml extracts showed the highest cytotoxicity with 83% against gastric carcinoma (AGS). The extracts showed weak antimicrobial activities against Bacillus subtilis and Pseudomonas aeruginosa, but practically no antimicrobial activity against the other microorganisms tested. The effect of ${\alpha}$-glucosidase inhibition was 64% at the concentration of 1.0 mg/ml. The inhibitory effect of angiotensin converting enzyme (ACE) of the extract in the range of $0.2{\sim}1.0mg/ml$ showed $63{\sim}89%$, and the highest ACE inhibition was 89% at the concentration of 1.0 mg/ml of extracts. The highest activity of glutathion S-transferase (GST) was 221 % at the concentration of 1.0 mg/ml of the G. elata extracts. These results suggest that G. elata may be used as health/functional food materials.