• Title/Summary/Keyword: Hsp40

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Heat Shock Causes Oxidative Stress and Induces a Variety of Cell Rescue Proteins in Saccharomyces cerevisiae KNU5377

  • Kim, Il-Sup;Moon, Hye-Youn;Yun, Hae-Sun;Jin, Ing-Nyol
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.492-501
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    • 2006
  • In this study, we attempted to characterize the physiological response to oxidative stress by heat shock in Saccharomyces cerevisiae KNU5377 (KNU5377) that ferments at a temperature of $40^{\circ}C$. The KNU5377 strain evidenced a very similar growth rate at $40^{\circ}C$ as was recorded under normal conditions. Unlike the laboratory strains of S. cerevisiae, the cell viability of KNU5377 was affected slightly under 2 hours of heat stress conditions at $43^{\circ}C$. KNU5377 evidenced a time-dependent increase in hydroperoxide levels, carbonyl contents, and malondialdehyde (MDA), which increased in the expression of a variety of cell rescue proteins containing Hsp104p, Ssap, Hsp30p, Sod1p, catalase, glutathione reductase, G6PDH, thioredoxin, thioredoxin peroxidase (Tsa1p), Adhp, Aldp, trehalose and glycogen at high temperature. Pma1/2p, Hsp90p and $H^+$-ATPase expression levels were reduced as the result of exposure to heat shock. With regard to cellular fatty acid composition, levels of unsaturated fatty acids (USFAs) were increased significantly at high temperatures ($43^{\circ}C$), and this was particularly true of oleic acid (C18:1). The results of this study indicated that oxidative stress as the result of heat shock may induce a more profound stimulation of trehalose, antioxidant enzymes, and heat shock proteins, as well as an increase in the USFAs ratios. This might contribute to cellular protective functions for the maintenance of cellular homeostasis, and may also contribute to membrane fluidity.

Bag-1L is a Stress-withstand Molecule Prevents the Downregulation of Mcl-1 and c-Raf Under Control of Heat Shock Proteins in Cisplatin Treated HeLa Cervix Cancer Cells

  • Ozfiliz, Pelin;Arisan, Elif Damla;Coker-Gurkan, Ajda;Obakan, Pinar;Eralp, Tugce Nur;Dinler-Doganay, Gizem;Palavan-Unsal, Narcin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.11
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    • pp.4475-4482
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    • 2014
  • Background: Cisplatin, a DNA damaging agent, induces apoptosis through increasing DNA fragmentation. However, identification of intrinsic resistance molecules against Cisplatin is vital to estimate the success of therapy. Bag-1 (Bcl-2-associated anthanogene) is one anti-apoptotic protein involved in drug resistance impacting on therapeutic efficiency. Elevated levels of this protein are related with increase cell proliferation rates, motility and also cancer development. For this reason, we aimed to understand the role of Bag-1 expression in Cisplatin-induced apoptosis in HeLa cervix cancer cells. Cisplatin decreased cell viability in time- and dose-dependent manner in wt and Bag-1L+HeLa cells. Although, $10{\mu}M$ Cisplatin treatment induced cell death within 24h by activating caspases in wt cells, Bag-1L stable transfection protected cells against Cisplatin treatment. To assess the potential protective role of Bag-1, we first checked the expression profile of interacting anti-apoptotic partners of Bag-1. We found that forced Bag-1L expression prevented Cisplatin-induced apoptosis through acting on Mcl-1 expression, which was reduced after Cisplatin treatment in wt HeLa cells. This mechanism was also supported by the regulation of heat shock protein (Hsp) family members, Hsp90 and Hsp40, which were involved in the regulation Bag-1 interactome including several anti-apoptotic Bcl-2 family members and c-Raf.

경구투여 백신 후보물질로서의 Helicobacter pylori 외막 단백질의 조사

  • 박형배;최태부
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.129-136
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    • 1997
  • Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

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The Assessment of Toxicity on organic Sludge Using Acetylcholinesterase, Cytochrome P450, and Hsp70 Extracted from Earthworm (Eisenia fetida) (지렁이에서 추출한 Acetylcholinesterase, Cytochrome P450, and Heat Shock protein 70을 이용한 유기성슬러지 독성 평가)

  • Na, Young-Eun;Bang, Hae-Son;Kim, Myung-Hyun;Kim, Min-Kyoung;Roh, Kee-An;Lee, Jung-Taek;Ahn, Young-Joon;Yoon, Seong-Tak
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.4
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    • pp.274-279
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    • 2007
  • The toxicitiy of organic sludge such as municipal sewage sludge (MSS), industrial sewage sludge (ISS), alcohol fermentation processing sludge (AFPS) and leather processing sludge (LPS) were evaluated with three environmental biomarkers as acetylcholinesterase, cytochrome P450, and heat shock protein 70 extracted from earthworm (Eisenia fetida). Their toxicities were compared with those of pig manure compost (PMC). MSS, ISS, LPS, and AFPS did not significantly affect the acetylcolinesterase activity, whereas only the elutriate of PMC slightly was increased the activity. MSS, AFPS, and PMC tended to slightly inhibit the cytochrome $P_{450}$ activity, but ISS and LPS showed significantly the inhibitory effect on cytochrome $P_{450}$. The hsp70 expression began to increase after treatments and showed high induction at 6 hour, followed by zero level at around 12 hour. The quantity of the hsp70 expressed by elutriate treatments of PMC, AFPS, MSS, ISS, and LPS was 1.9, 3.0, 3.3, 4.4, and 4.7 fold higher than that of distilled water. These results indicate that in toxicity tests of five organic waste materials, four kinds of sludge materials appeared more toxic than PMC. Results of AChE, P450, and hsp70 of earthworm might be useful for expecting or assessing an effect by exposure of organic wastes to earthworms in soil.

Eukaryotic DNAJ/K Database: A Comprehensive Phylogenomic Analysis Platform for the DNAJ/K Family

  • Cheong, Kyeongchae;Choi, Jaehyuk;Choi, Jaeyoung;Park, Jongsun;Jang, Suwang;Lee, Yong-Hwan
    • Genomics & Informatics
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    • v.11 no.1
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    • pp.52-54
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    • 2013
  • Proteins in DNAJ/K families are ubiquitous, from prokaryotes to eukaryotes, and function as molecular chaperones. For systematic phylogenomics of the DNAJ/K families, we developed the Eukaryotic DNAJ/K Database (EDD). A total of 12,908 DNAJs and 4,886 DNAKs were identified from 339 eukaryotic genomes in the EDD. Kingdom-wide comparison of DNAJ/K families provides new insights on the evolutionary relationship within these families. Empowered by 'class', 'cluster', and 'taxonomy' browsers and the 'favorite' function, the EDD provides a versatile platform for comparative genomic analyses of DNAJ/K families.

The Effects of Dietary Supplementation of Vitamin C or E on the Expressions of Endoplasmic Reticulum Stress, Lipid and Glucose Metabolism Associated Genes in Broiler Chickens (비타민 C 및 E의 첨가 급여가 육계의 소포체 스트레스와 지방 및 포도당 대사 연관 유전자의 발현에 미치는 영향)

  • Park, Jeong Geun;An, Young Sook;Sohn, Sea Hwan;Jang, In Surk;Moon, Yang Soo
    • Korean Journal of Poultry Science
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    • v.40 no.2
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    • pp.147-155
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    • 2013
  • This study was conducted to evaluate the effects of dietary supplementation of vitamin C or E on the expressions of endoplasmic reticulum (ER) stress, lipid and glucose metabolism associated genes in broiler chickens. A total of 216 one-day-old male broilers was randomly alloted to 4 treatments with 6 replicate pens per treatment and 9 broilers per pen for 35 days. The dietary treatments were control, vitamin C (control diet + ascorbic acid 200 mg/kg diet), vitamin E (control diet + ${\alpha}$-tocopherol 100 mg/kg diet), vitamin C + E (control diet + vitamin C 200 mg/kg + vitamin E 100 mg/kg), respectively. To evaluate gene expressions by quantitative real-time polymerase chain reaction, total RNA was extracted from the liver of the chicken at 35 days of age. Dietary supplementation of vitamins was significantly down-regulated the expression of stress marker genes including HSP70, HSP90, and HMGCR, as compared to the control (p<0.05). The expressions of ER stress associated genes also inhibited by supplementation of vitamins as well (p<0.05). Vitamin C supplementation suppressed the expression of lipid associated genes such as FASN, FATP1 and ACSL1. Vitamin supplementation did not affect the glucose transporters, GLUT2 and GLUT8, in the liver. The results of the present study indicated that dietary supplementation of vitamin C or E could be beneficial for the alleviating physiological stress in broiler chickens.

Classification of Biological Effect of 1,763 MHz Radiofrequency Radiation Based on Gene Expression Profiles

  • Im, Chang-Nim;Kim, Eun-Hye;Park, Ae-Kyung;Park, Woong-Yang
    • Genomics & Informatics
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    • v.8 no.1
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    • pp.34-40
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    • 2010
  • Radiofrequency (RF) radiation might induce the transcription of a certain set of genes as other physical stresses like ionizing radiation and UV. To observe transcriptional changes upon RF radiation, we exposed WI-38, human lung fibroblast cell to 1763 MHz of mobile phone RF radiation at 60 W/kg of specific absorption rate (SAR) for 24h with or without heat control. There were no significant changes in cell numbers and morphology after exposure to RF radiation. Using quantitative RT-PCR, we checked the expression of three heat shock protein (HSP) (HSPA1A, HSPA6 and HSP105) and seven stress-related genes (TNFRSF11B, FGF2, TGFB2, ITGA2, BRIP1, EXO1, and MCM10) in RF only and RF/HS groups of RF-exposed cells. The expressions of three heat shock proteins and seven stress-related genes were selectively changed only in RF/HS groups. Based on the expression of ten genes, we could classify thermal and non-thermal effect of RF-exposure, which genes can be used as biomarkers for RF radiation exposure.

Effects of Lipopolysaccride-induced Stressor on the Expression of Stress-related Genes in Two Breeds of Chickens (Lipopolysaccride 감염처리가 닭의 품종간 스트레스연관 유전자 발현에 미치는 영향)

  • Jang, In Surk;Sohn, Sea Hwan;Moon, Yang Soo
    • Korean Journal of Poultry Science
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    • v.44 no.1
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    • pp.1-9
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    • 2017
  • The objective of the present study was to determine the expression of genes associated with lipopolysaccharide (LPS)-induced stressor in two breeds of chickens: the Korean native chicken (KNC) and the White Leghorn chicken (WLH). Forty chickens per breed, aged 40 weeks, were randomly allotted to the control (CON, administered the saline vehicle) and LPS-injected stress groups. Samples were collected at 0 and 48 h post-LPS injection, and total RNA was extracted from the chicken livers for RNA microarray and quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In response to LPS, 1,044 and 1,193 genes were upregulated, and 1,000 and 1,072 genes were downregulated in the KNC and WLH, respectively, using a ${\geq}2$-fold cutoff change. A functional network analysis revealed that stress-related genes were downregulated in both KNC and WLH after LPS infection. The results obtained from the qRT-PCR analysis of mRNA expression of heat shock 90 (HSP90), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), activating transcription factor 4 (ATF4), sterol regulatory element-binding protein 1 (SREBP1), and X-box binding protein 1 (XBP1) were confirmed by the results of the microarray analysis. There was a significant difference in the expression of stress-associated genes between the control and LPS-injected KNC and WLH groups. The qRT-PCR analysis revealed that the stress-related $HSP90{\alpha}$ and HMGCR genes were downregulated in both LPS-injected KNC and WLH groups. However, the HSP70 and $HSP90{\beta}$ genes were upregulated only in the LPS-injected KNC group. The results suggest that the mRNA expression of stress-related genes is differentially affected by LPS stimulation, and some of the responses varied with the chicken breed. A better understanding of the LPS-induced infective stressors in chicken using the qRT-PCR and RNA microarray analyses may contribute to improving animal welfare and husbandry practices.

Stress Evaluation to Heavy Metal Exposure using Molecular Marker in Chironomus riparius (분자지표 유전자 발현을 통한 Chironomus riparius 중금속 노출 스트레스 평가)

  • Kim, Won-Seok;Park, Kiyun;Kwak, Ihn-Sil
    • Korean Journal of Ecology and Environment
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    • v.53 no.2
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    • pp.165-172
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    • 2020
  • Heavy metals are common pollutants in the freshwater environment and have toxicological effect in habitat organisms. The heavy metals highly accumulated in sediment and organism, and observed various physiological responses. In this study, we investigated the molecular response to heavy metal toxicity (Al, Aluminum; Cr, Chromium; Cu, copper; Mn, Manganese; Zn, Zinc) through expression of heat shock protein 40, 70, 90 (HSP40, 70, 90), cytochrome 450 (CYP450), Glutathione S-transferase (GST) and Serine-type endopeptidase (SP). HSPs showed up-regulation in Cu and Zn exposures. Furthermore, HSPs expression in treated groups tended to be higher than the control group. The tendency of CYP450 and GST mRNA expression was higher for Cr and Cu than for other exposure group. The expression of SP gene was low at Al exposure and other group were measured to be similar to control. These results suggest that heavy metal toxicity in freshwater ecosystem may affect physiological and molecular process. Also, the comprehensive gene expression in the aquatic midge Chironomus riparius give useful information to potential molecular biomarkers for assessing heavy metal toxicity.

Effects of pre-heat treatments on the quality of cut kimchi cabbages during short-term storage (절단배추의 단기 저장에 대한 예열처리 효과)

  • Kim, Sang-Seop;Ku, Kyung-Hyung;Jeong, Moon-Cheol;Hong, Joo-Heon;Chung, Shin-Kyo
    • Food Science and Preservation
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    • v.21 no.6
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    • pp.776-783
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    • 2014
  • Kimchi cabbages were cut ($3{\times}3cm$), and were pre-heat treated at $40^{\circ}C$, and their physicochemical qualities and browning degrees were investigated during 8 weeks storage at $5^{\circ}C$. The Cut kimchi cabbages were treated at $40^{\circ}C$ (1~8 hrs) and their protein bands profiles were determined by sodium dodecyl sulfate polyacrylamide electrophoresis (SDS-PAGE). The 60, 39, 33, and 12 kDa bands considered heat shock proteins (HSPs) were expressed in the cut kimchi cabbage, and the 4-hr pre-heat treatment (HS 4) exhibited the strongest band ratio. The weight ratios and titratable acidities of the pre-heat treated cut kimchi cabbages were not changed so much after 8 weeks storage at $5^{\circ}C$, and the soluble solid contents of HS 4 decreased less than that of any other treatments. The browning degree of HS 4 after 8 week storage was also shown to be the least among the treatments. The polyphenol oxidase (PPO) activities of all treatments slightly rose during the over all storage period, in contrast with the decrease of total phenolic contents. The expression of HSPs was identified in the pre-heat treated cut kimchi cabbages, and HS 4 exhibited the best quality and appearance after 8 weeks storage at $5^{\circ}C$.