Kim, Min-Gab;Su'udi, Mukhamad;Park, Sang-Ryeol;Hwang, Duk-Ju;Bae, Shin-Chul
Journal of Life Science
/
v.20
no.12
/
pp.1777-1783
/
2010
Plants generate reactive oxygen species (ROS) as a by-product of normal aerobic metabolism or when exposed to a variety of stress conditions, which can cause widespread damage to biological macromolecules. To protect themselves from oxidative stress, plant cells are equipped with a wide range of antioxidant proteins. However, the detailed reaction mechanisms of these are still unknown. Peroxiredoxins (Prxs) are ubiquitous thiol-containing antioxidants that reduce hydrogen peroxide with an N-terminal cysteine. The active-site cysteine of peroxiredoxins is selectively oxidized to cysteine sulfinic acid during catalysis, which leads to inactivation of peroxidase activity. This oxidation was thought to be irreversible. Recently identified small protein sulphiredoxin (Srx1), which is conserved in higher eukaryotes, reduces cysteine.sulphinic acid in yeast peroxiredoxin. Srx1 is highly induced by $H_2O_2$-treatment and the deletion of its gene causes decreased yeast tolerance to $H_2O_2$, which suggest its involvement in the metabolism of oxidants. Moreover, Srx1 is required for heat shock and oxidative stress induced functional, as well as conformational switch of yeast cytosolic peroxiredoxins. This change enhances protein stability and peroxidase activity, indicating that Srx1 plays a crucial role in peroxiredoxin stability and its regulation mechanism. Thus, the understanding of the molecular basis of Srx1 and its regulation is critical for revealing the mechanism of peroxiredoxin action. We postulate here that Srx1 is involved in dealing with oxidative stress via controlling peroxiredoxin recycling in Arabidopsis. This review article thus will be describing the functions of Prxs and Srx in Arabidopsis thaliana. There will be a special focus on the possible role of Srx1 in interacting with and reducing hyperoxidized Cys-sulphenic acid of Prxs.
Parkinson's disease (PD) is a common neurodegenerative disorder and is characterized by the progressive loss of dopaminergic neurons in the substantia nigra. Although many studies showed that the aggregation of $\alpha$-synuclein might be involved in the pathogenesis of PD, its protective properties against oxidative stress remain to be elucidated. In this study, human wild type and mutant $\alpha$-synuclein genes were fused with a gene fragment encoding the nine amino acid trans activator of transcription (Tat) protein transduction domain of HIV-l in a bacterial expression vector to produce a genetic in-frame WT Tat-$\alpha$-synuclein (wild type) and mutant Tat-a-synucleins (mutants; A30P and A53T), respectively, and we investigated the protective effects of wild type and mutant Tat-$\alpha$-synucleins in vitro and in vivo. WT Tat-$\alpha$-synuclein rapidly transduced into an astrocyte cells and protected the cells against paraquat induced cell death. However, mutant Tat-$\alpha$-synucleins did not protect at all. In the mice models exposed to the herbicide paraquat, the WT Tat-$\alpha$-synuclein completely protected against dopaminergic neuronal cell death, whereas mutants failed in protecting against oxidative stress. We found that these protective effects were characterized by increasing the expression level of heat shock protein 70 (HSP70) in the neuronal cells and this expression level was dependent on the concentration of transduced WT Tat-$\alpha$-synuclein. These results suggest that transduced Tat-$\alpha$-synuclein might protect cell death from oxidative stress by increasing the expression level of HSP70 in vitro and in vivo and this may be of potential therapeutic benefit in the pathogenesis of PD.
Garlic occupies a special position among the many foods of vegetable origin because it is the sole food for Koreans during the their lives. And vitamin A has been ingested by forms of food or additives. Cadmium has been described as one of the most dangerous trace elements in the food and environment of man and livestocks. Since the de novo synthesis of stress proteins can be detected early after exposure to some agents, analysis of cadmium-induced changes in gene expression , ie. alterations in patterns of protein synthesis, may be useful to develop as biomarkers of exposure and damage for food hygiene. He acute and chronic combine effects of cadmium (Cd, CdCl2 20mg/kg), garlic oil(Dds: diallyl disulfide 50mg/kg, 3 times a week) and vitamin A(Ra: retinol acetate 50,000 IU/kg, 3 times a week) on Wistar male rats were evaluated concerning cadmium contents, tissues enzyme activity, HSP expression histopathological and electron microscopical examinations. The results of the study are as follows ; 1. Less cadmium was absorbed through the digestive tracts, but the ratio of contents in tissue were not changed by the simultaneous adminstration of diallyl disufide or retinol acetate. 2. ALT(alanine aminotransferase) , AST(aspartate aminotransferase), glucose, BUN (blood urea nitrogen), creatinine, the key indices of the clinical changes in hepatic and renal function were significantly hanged by the cadmium treatment after 1 week in liver, after 4 weeks in kidney. 3. Histopathological changes in cadmium treated rats were appeared at 8 weeks age treatment in kidneys. Homogenous eosinophilic material was accumulated in cortical and collecting tubular lumens at 16 weeks. Degenerated or necrotized tubular cells were observed in cortex and medulla. Degenerated seminiferous tubules and homogeneous eosinophilic material was seen in interstitial tissue of rat treated with cadmium for 16 weeks. Calcium deposits were seen in degenerated seminiferous tubules and the tubules showed severe calcification of rat treated with cadmium for 16 weeks. Electron microscope changes in kidney were observed in rats treated with CdCl2 20 mg/kg. Proximal convoluted tubule cells showed selling of cytoplasm and narrow lumen. Capillary endothelial cells showed cytoplasmic vacuoles and swelling. Degenerated epithelial cells were accumulated in tubular lumen of kidney. 4. Enhanced synthesis of 70 KDa relateve molecular mass proteins were detected in 2 hours after cadmium, exposure, with maximum activity occurring at 8~48 hours. Induction of HSP 70 was evident at proximal tubules and glomeruli in kidney. Testicular cells produced enough HSP to be detected normally. From the above results, it could be concluded that HSP70 induction by the cadmium treatment was a rapid reaction to indicated the exposure of xenobiotics, and retinol acetate reduced the cadmium induced nephrotoxicity.
Human epidermal growth factor receptor 2 (HER2) inhibitors, such as trastuzumab and lapatinib are used to treat HER2-positive breast and gastric cancers. However, as with other targeted therapies, intrinsic or acquired resistance to HER2 inhibitors presents unresolved therapeutic problems for HER2-positive gastric cancer. The present study describes investigations with AUY922, a heat shock protein 90 (HSP90) inhibitor, in primary lapatinib-resistant (ESO26 and OE33) and lapatinib-sensitive gastric cancer cells (OE19, N87, and SNU-216) harboring HER2 amplification/over-expression. In order to investigate whether AUY922 could overcome intrinsic and acquired resistance to HER2 inhibitors in HER2-positive gastric cancer, we generated lapatinib-resistant gastric cancer cell lines (OE19/LR and N87/LR) by continuous exposure to lapatinib in vitro. We found that activation of HER2 and protein kinase B (AKT) were key factors in inducing intrinsic and acquired lapatinib-resistant gastric cancer cell lines, and that AUY922 effectively suppressed activation of both HER2 and AKT in acquired lapatinib-resistant gastric cancer cell lines. In conclusion, AUY922 showed a synergistic anti-cancer effect with lapatinib and sensitized gastric cancer cells with intrinsic resistance to lapatinib. Dual inhibition of the HSP90 and HER2 signaling pathways could represent a potent therapeutic strategy to treat HER2-positive gastric cancer with intrinsic and acquired resistance to lapatinib.
Jong Woo Park;Chang Hoon Lee;Chan Young Jeong;Hyeok Gyu Kwon;Seul Ki Park;Ji Hae Lee;Sang Kuk Kang;Seong-Wan Kim;Seong-Ryul Kim;Hyun-Bok Kim;Kee Young Kim
International Journal of Industrial Entomology and Biomaterials
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v.46
no.1
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pp.16-23
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2023
The silkworm's dormancy and embryonic development are accomplished through the interaction of various genes. Analysis of the expression of several interacting genes can predict the embryonic stage of silkworms. In this study, we analyzed the changes in the expression level of genes at each stage during the embryonic development of dormant silkworm eggs and selected genes that can predict the hatching time. Jam123 and Jam124 silkworms were collected after egg laying, and the silkworm eggs were preserved using a double refrigeration method and expression analysis was performed for 23 genes during embryogenesis. There were 5 genes showing significant changes during embryogenesis: UDP-glucuronosyltransferases (BmUGTs), heat shock protein hsp20.8 (BmHsp20.8), Cytochromes b5-like proteins (BmCytb5), Krüppel homolog 1 (BmKr-h1), and cuticular protein RR-1 motif 41 (BmCpr41). As a result of quantitative comparison of the expression levels of these 5 genes through real-time PCR, the BmUGTs gene showed a difference between Jam123 and Jam124, making it difficult to see it as an indicator for predicting hatching time. However, the BmHsp20.8 gene had a common expression decreased at the imminent hatching stage. In addition, it was confirmed that the expression level of the BmCytb5 gene decreased to the lowest level at the time of imminent hatching, and the expression of the BmKr-h gene was made only at the time of imminent hatching. The expression of the last BmCpr41 gene can be confirmed only at the time of imminent hatching, and it was confirmed that it shows a rapid increase right before hatching. Taken together, these results suggest that expression analysis of BmHsp20.8, BmCytb5, BmKr-h1, and BmCpr41 genes can determine the stage of embryogenesis, predict hatching time, which facilitate better management of silkworm eggs.
Differentially Expressed Gene (DEG) was obtained from Differential Display Reverse Transcription (DDRT)-PCR using Annealing Control Primer (ACP) to search and clone genes related to developmental stages of Sebastes inermis. By using 120 ACPs, the nucleotide sequences obtained from 16 DEGs showing higher expression in 6-month-old skeletal muscle than 18-month-old ones and from 22 DEGs displaying stronger expression in 18-month-old than 6-month-old were analyzed and BLAST was conducted. The results identified that DEGs shared 69~95% homology with genes of parvalbumin (PVALB), nucleoside diphosphate kinase (NDK) B, tropomyosin (TPM), troponin I (TnI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), muscle-type creatine kinase (CKM2), small EDRK-rich factor 2 (SERF2), adenosine monophosphate deaminase (AMPD), Trimeric intracellular cation channel type A (TRICA), Rho GTPase-activating protein 15 (ARHGAP15), S-formylglutathione hydrolase (Esterase D; ESD), heat shock protein 70 (hsp70), type 1 collagen alpha 2 (COL1A2), glutathione S-transferase, Mid1-interacting protein 1 (Mid1lip1), myosin light chain 1 (MYL1), sarcoplasmic/endoplasmic reticulum calcium ATPase 1B (SERCA1B), and ferritin heavy subunit (FTH1). Expression pattern by developmental stage of DEG14 and PVALB exhibiting strong expression in 6-month-old skeletal muscle was investigated using real time PCR. Expression was reduced as Sebastes inermis grew. Expression of PVALB gene was extremely low after 6 months of age. Expression of CKM2 showed higher expression in 18-month-old skeletal muscle than in 6-month-old muscles, and increased continuously until 4 years old, after which CKM2 expression became gradually reduced. By analysis of tissue-specific expression patterns of DEG, DEG14 was expressed mainly in skeletal muscle, liver, kidney and spleen tissues, whereas PVALB expression was expressed in skeletal muscle and kidney, but not in liver and spleen tissues. CKM2 was expressed in skeletal muscle, kidney, and spleen tissues, but not in liver tissues. PVALB gene was composed of 110 amino acids, which constituted 659 bp nucleotides. The results reported here demonstrate that the expression patterns of parvalbumin and CKM2 could be used as molecular markers for selecting fishes exhibiting fast growth.
Park, Yu Kyung;Lee, Chang-Eun;Lee, Hyoungseok;Koh, Hye Yeon;Kim, Sojin;Lee, Sung Gu;Kim, Jung Eun;Yim, Joung Han;Hong, Ju-Mi;Kim, Ryeo-Ok;Han, Se Jong;Kim, Il-Chan
Journal of Life Science
/
v.30
no.11
/
pp.931-938
/
2020
Translationally controlled tumor protein (TCTP) is one of the most abundant proteins in various eukaryotic organisms. TCTPs play important roles in cell physiological processes in cancer, cell proliferation, gene regulation, and heat shock response. TCTP is also considered an important factor in the resistance to oxidative stress induced by dithiothreitol or hydrogen peroxide (H2O2). Arctic calanoid copepods have a variety of antioxidant defense systems to regulate the levels of potentially harmful reactive oxygen species generated by ultraviolet radiation in the Arctic marine ecosystem. However, information on the antioxidant activity of TCTP in the Arctic Calanus glacialis is still scarce. To understand the putative antioxidant function of the Arctic copepod C. glacialis TCTP (Cg-TCTP), its gene was cloned and sequenced. The Cg-TCTP comprised 522 bp and encoded a 174-amino acid putative protein with a calculated molecular weight of ~23 kDa. The recombinant Cg-TCTP (Cg-r TCTP) gene was overexpressed in Escherichia coli (BL21), and Cg-rTCTP-transformed cells were grown in the presence or absence of H2O2. Cg-rTCTP-transformed E. coli showed increased tolerance to high H2O2 concentrations. Therefore, TCTP may be an important antioxidant protein related to tolerance of the Arctic copepod C. glacialis to oxidative stress in the harsh environment of the Arctic Ocean.
Sohn, Sea Hwan;Cho, Eun Jung;Park, Ji Ae;Hong, Young Ho;Kim, Chong Dae
Korean Journal of Poultry Science
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v.42
no.2
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pp.157-167
/
2015
We compared the degrees of stress response of 12 domestic purebred chicken strains that have been bred at National Institute of Animal Science, RDA, Korea since 1980. As a physiological marker of stress response, the expression levels of heat shock protein (HSP)-70, HSP-$90{\alpha}$, HSP-$90{\beta}$, hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) genes and telomere length were measured by quantitative real-time polymerase chain reaction using the lymphocytes of 1,101 chickens. There was significant difference in HSP-70, HSP-$90{\alpha}$, HMGCR expression and telomere length among the strains. There was also significant difference in HSP-$90{\alpha}$, HSP-$90{\beta}$, and HMGCR expression between male and female chickens. Different age groups of chicken exhibited different expression levels of HSP-70, HSP-$90{\alpha}$ and telomere length. The results of the HSPs expression level suggested that, the strains of R, L and Y were highly resistant to stress, whereas the strains of S, O and W were susceptible to stress. Although the statistical differences in some of HSPs gene expression existed between genders, the HSP expression results varied in different strains that some opposed to the others, and there might be interaction between strains and genders, which conclude that there was no difference in stress response between male and female chickens. Moreover, despite of significant difference in some of HSPs expression level, it was considered that there was no difference in stress response between ages due to the inconsistent trends among HSP markers.
This study investigated the effects of feeds naturally contaminated with mycotoxins on growth performance, serum biochemical parameters, carcass traits, and splenic heat shock protein 70 (Hsp70) mRNA expression levels in broiler chickens. The efficacy of yeast cell wall (YCW) adsorbent in preventing mycotoxicosis was also evaluated. Three hundred 1-d-old Arbor Acres broiler chicks were randomly allotted to 3 treatments in completely randomized design for 42 d. Each treatment group had 5 replicate pens with 20 birds. The treatments were as follows: i) basal diet (control), ii) naturally contaminated diet (NCD), and iii) NCD+0.2% YCW adsorbent (NCDD). The NCD decreased average daily gain (ADG) (p<0.01) of 0 to 21 d, 22 to 42 d, and 0 to 42 d, and increased feed conversion ratio (p<0.01) of 22 to 42 d and 0 to 42 d. Both the breast meat percentage and thigh meat percentage of the NCD group were significantly higher (p<0.01) than that of the control group on d 21. The NCD group showed significantly increased levels of triglycerides (p<0.05) and cholesterol (p<0.05) on both d 21 and d 42 compared to the control group. However, the NCD significantly reduced (p<0.01) the high-density lipoprotein (HDL) on d 42 compared to controls. Compared with the NCD, supplementation with YCW significantly improved (p<0.01) the ADG of 0 to 21 d and 0 to 42 d, and increased (p<0.01) concentrations of HDL on d 42, and on d 21, and triglycerides (p<0.05) on d 21 and d 42. Supplementation with YCW reduced (p<0.01) the breast meat percentage, the thigh meat percentage, the concentrations of cholesterol (p<0.01) and the low-density lipoprotein (p<0.05) on d 21, and improved (p<0.01) the splenic Hsp70 mRNA expression levels compared with the NCD group. The results of this study indicated that feeding NCD for 42 d had adverse effects on broiler chickens, and that YCW might be beneficial in counteracting the effects of mycotoxins.
Kim Tae-Hyung;Yang Moon-Hyu;Choe Mi-Kyung;Han Seok-Jung;Yeo In-Kyu
Journal of Aquaculture
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v.18
no.1
/
pp.7-12
/
2005
This study was conducted to investigate antioxidant enzyme activity (catalase and superoxide dismutase) and Heat Shock Protein 70 (HSP70) mRNA variation in hepatopancreas of abalone (Haliotis discus hannai) cultured under several acute water temperatures. Abalones were cultured at 10, 15, 20, 25 and $30^{\circ}C$, for 0, 6, 12, 24 and 48 hours, respectively. The HSP70 mRNA expression in hepatopancreas was more increased at $30^{\circ}C$ compared to those at 10. 15, 20 (control) and $25^{\circ}C$. The superoxide dismutase (SOD) activity was increased in hepato-pancreas at all water temperature conditions compared to the control ($20^{\circ}C$). The SOD activity at high water temperature (25 and $30^{\circ}C$) tended to be increased after 12 hours, and was increased immediately after exposure to low water temperature (10 and $15^{\circ}C$). and then was recovered to starting level after the increase. Also, catalase (CAT) activity in hepatopancreas was increased in all the groups except for at $10^{\circ}C$ than the control ($20^{\circ}C$). Survival rate of abalone was $100\%$ at 10, 15, 20 and $25^{\circ}C$, but $92\%$ at $30^{\circ}C$. Thus, according to our study, when abalone is appeared at $20^{\circ}C$, defense mechanism against stress at low water temperature can be accelerated to be stabilized at about $5^{\circ}C$. In the case of exposure of abalone to high water temperature, antioxidant enzyme and HSP70 expression were increased due to elevated physiological stimulation factor, such as temperature.
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