• Title/Summary/Keyword: Heat shock factor-1

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Search for Novel Stress-responsive Protein Components Using a Yeast Mutant Lacking Two Cytosolic Hsp70 Genes, SSA1 and SSA2

  • Matsumoto, Rena;Rakwal, Randeep;Agrawal, Ganesh Kumar;Jung, Young-Ho;Jwa, Nam-Soo;Yonekura, Masami;Iwahashi, Hitoshi;Akama, Kuniko
    • Molecules and Cells
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    • v.21 no.3
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    • pp.381-388
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    • 2006
  • Heat shock proteins (Hsp) 70 are a ubiquitous family of molecular chaperones involved in many cellular processes. A yeast strain, ssa1/2, with two functionally redundant cytosolic Hsp70s (SSA1 and SSA2) deleted shows thermotolerance comparable to mildly heatshocked wild type yeast, as well as increased protein synthesis and ubiquitin-proteasome protein degradation. Since mRNA abundance does not always correlate well with protein expression levels it is essential to study proteins directly. We used a gel-based approach to identify stress-responsive proteins in the ssa1/2 mutant and identified 43 differentially expressed spots. These were trypsin-digested and analyzed by nano electrospray ionization liquid chromatography tandem mass spectrometry (nESI-LC-MS/MS). A total of 22 non-redundant proteins were identified, 11 of which were confirmed by N-terminal sequencing. Nine proteins, most of which were up-regulated (2-fold or more) in the ssa1/2 mutant, proved to be stress-inducible proteins such as molecular chaperones and anti-oxidant proteins, or proteins related to carbohydrate metabolism. Interestingly, a translational factor Hyp2p up-regulated in the mutant was also found to be highly phosphorylated. These results indicate that the cytosolic Hsp70s, Ssa1p and Ssa2p, regulate an abundance of proteins mainly involved in stress responses and protein synthesis.

Putative response regulator two-component gene, CaSKN7, regulate differentiation and virulence in Candida albicans

  • Lee, Jung-Shin;Minyoung Lim;Yim, Hyung-Soon;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.50-50
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    • 2003
  • We have identified and analysed a putative response regulator two-component gene (CaSKN7) from Candida albicans and its encoding protein (CaSkn7). CaSKN7 has an open reading frame of 1677bp. CaSKN7 encodes a 559 amino acid protein (CaSkn7) with an estimated molecular mass of 61.1 kDa. CaSKN7 is a homologue of a Saccharomyces cerevisiae SKN7 that is the regulator involved in the oxidative stress response. To study the role of CaSKN7, we constructed a CAI4-derived mutant strain carrying a homozygous deletion of the CaSKN7 gene. In the caskn7 disruptant cells, the formation of germ tube require shorter time than that in the congenic wild-type strain but the growth of mycelium delayed in liquid media. In contrast, the caskn7 disruptant cells attenuate the differentiation in solid media and the virulence in mouse model system. Expression level of hypha-specific and virulence genes - HYR1, ECE1, HWP1, and ALS1 - in the caskn7 disruptant cells increased as compared with that in the congenic wild-type strain in 10% serum YPD. Skn7 in 5. cerevisiae was found to bind the HSE element from the SSA promoter, Also, CaSkn7 contains heat shock factor DNA-binding domain and the promoters of these genes have HSE-like sties. Therefore these results show that CaSKN7 regulate the differentiation and virulence of C. albicans.

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Inhibitory Effect of Acute Pancreatitis in Rats by Patrinia Scabiosaefolia (급성췌장염 유발된 흰쥐에 대한 패장의 억제 효과)

  • Lee, Joon-Suk;Jung, Sang-Pil;Kil, Eun-Young;Lee, Su-Kyung;Kwon, Young-Dal;Song, Yung-Sun
    • Journal of Korean Medicine Rehabilitation
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    • v.15 no.1
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    • pp.99-108
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    • 2005
  • Objectives : Patrinia scabiosaefolia (PS) has long been as a remedy for treating infectious diseases in Korea. In the present experiments, the author examined the effects of PS on the cholecystokinin-octapeptide (CCK)-induced pancreatitis (AP) in rats. Methods : Male Wister rats weighing 200 to 250 g were divided into two group. Normal untreated group, in treatment with PS group; PS was administered orally, followed by $75{\mu}g/kg$ CCK subcutaneously three times, after 1, 3 and 5 h. This whole procedure was repeated for 5 days. In treatment with saline group, the protocol was the same as in treatment group with PS. The author determined the pancreatic weight/body weight ratio, the levels of pancreatic heat shock proteins(HSP)60, HSP72 and the secretion of pro-inflammatory cytokines. Results and Conclusion : PS was significantly decreased the pancreatic weight/body weight ratio in CCK-induced AP. PS increased HSP60 and HSP72 compared with CCK-induced AP. Additionally, the secretion of tumor necrosis factor(TNF)-${\alpha}$, interleukin(IL)-$1{\beta}$ and IL-6 the levels of amylase and lipase were lower than that of saline. These results suggest that PS may has a inhibitory effect against CCK-induced AP.

Proteomics Analysis of Gastric Epithelial AGS Cells Infected with Epstein-Barr Virus

  • Ding, Yong;Li, Xiao-Rong;Yang, Kai-Yan;Huang, Li-Hua;Hu, Gui;Gao, Kai
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.367-372
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    • 2013
  • Effects of the Epstein-Barr virus (EBV) on cellular protein expression are essential for viral pathogenesis. To characterize the cellular response to EBV infection, differential proteomes of gastric epithelial AGS cells were analyzed with two-dimensional gel electrophoresis (2-DE) followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and liquid chromatography electrospray/ionization ion trap (LC-ESI-IT) mass spectrometry identification. Mass spectrometry identified 9 altered cellular proteins, including 5 up-regulated and 4 down-regulated proteins after EBV infection. Notably 2-DE analysis revealed that EBV infection induced increased expression of heat shock cognate 71 kDa protein, actin cytoplasmic 1, pyridoxine-5'-phosphate oxidase, caspase 9, and t-complex protein 1 subunit alpha. In addition, EBV infection considerably suppressed those cellular proteins of zinc finger protein 2, cyclin-dependent kinase 2, macrophage-capping protein, and growth/differentiation factor 11. Furthermore, the differential expressional levels of partial proteins (cyclin-dependent kinase 2 and caspase 9) were confirmed by Western blot analysis.Thus, this work effectively provided useful protein-related information to facilitate further investigation of the mechanisms underlying EBV infection and pathogenesis.

Therapeutic Effects of Electroacupuncture on Cholecystokinin-octapeptide-induced Acute Pancreatitis Models (급성 췌장염모델에서 전침의 치료효과)

  • Cheong, Sang-Su;Yoon, Ji-Won;Jeong, Kyoung-Ah;Lee, Jong-Deok;Bai, Sun-Joon;Cho, Zang-Hee;Sung, Kang-Kyung
    • Journal of Acupuncture Research
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    • v.22 no.5
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    • pp.57-66
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    • 2005
  • Objectives : We examined the effects of electroacupuncture on the cholecystokinin-octapeptide-induced acute pancreatitis in rats. Methods : Rats were administered with $75{\mu}g/kg$ cholecystokinin-octapeptide subcutaneously three times (1, 3 and 5h after shaving) for 5days. Three days after finishing cholecystokinin-octapeptide administration, high frequency electroacupuncture (100Hz) and low frequency electroacupuncture (2Hz) were applied to acupoint equivalent to ST36 (Zusanli) for 7 days. The author determined the pancreatic weight/body weight ratio, the levels of pancreatic heat shock protein HSP60 and HSP72. The author also assay the secretion of ${\beta}-amylase$, lipase and pro-inflammatory cytokines in serum. Repeated cholecysokinin-octapeptide treatment resulted in the typical laboratory and morphological changes of experimentally induced pancreatitis. Results : Eelectroacupuncture significantly decreased the pancreatic weight/body weight ratio in cholecystokinin-octapeptide-induced acute pancreatitis, increased the pancreatic levels of HSP60 and HSP72, and decreased ${\beta}-amylase$ and lipase levels in cholecystokinin-octapeptide-induced acute pancreatitis. Additionally, the secretion of $Interleukin-1{\beta}$ and tumor necrosis $factor-{\alpha}$ was decreased in the animals treated with electroacupuncture. Conclusion : These results suggest that electroacupuncture may have protective effects against cholecystokinin-octapeptide-induced acute pancreatitis.

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Nitric Oxide as a Pro-apoptotic as well as Anti-apoptotic Modulator

  • Choi, Byung-Min;Pae, Hyun-Ock;Jang, Seon-Il;Kim, Young-Myeong;Chung, Hun-Taeg
    • BMB Reports
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    • v.35 no.1
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    • pp.116-126
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    • 2002
  • Nitric oxide (NO), synthesized from L-arginine by NO synthases, is a small, lipophilic, diffusible, highly reactive molecule with dichotomous regulatory roles in many biological events under physiological and pathological conditions. NO can promote apoptosis (pro-apoptosis) in some cells, whereas it inhibits apoptosis (anti-apoptosis) in other cells. This complexity is a consequence of the rate of NO production and the interaction with biological molecules such as metal ion, thiol, protein tyrosine, and reactive oxygen species. Long-lasting overproduction of NO acts as a pro-apoptotic modulator, activating caspase family proteases through the release of mitochondrial cytochrome c into cytosol, up-regulation of the p53 expression, and alterations in the expression of apoptosis-associated proteins, including the Bcl-2 family. However, low or physiological concentrations of NO prevent cells from apoptosis that is induced by the trophic factor withdrawal, Fas, $TNF{\alpha}$/ActD, and LPS. The anti-apoptotic mechanism is understood on the basis of gene transcription of protective proteins. These include: heat shock protein, hemeoxygenase, or cyclooxygenase-2 and direct inhibition of the apoptotic executive effectors caspase family protease by S-nitrosylation of the cysteine thiol group in their catalytic site in a cell specific way. Our current understanding of the mechanisms by which NO exerts both pro- and anti-apototic action is discussed in this review article.

Therapeutic effect of the traditional herbal formula, Hwanglyeonhaedok-tang, on rhinitis: A Review of the experimental study (황련해독탕이 비염 개선에 미치는 효과: 기초실험 연구 문헌 분석)

  • Son, Mi-Ju;Jerng, Ui-Min;Kim, Yun-Hee;Lee, Dong-Hyo;Kim, Sung-Ha
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.30 no.1
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    • pp.74-86
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    • 2017
  • Objectives : This study aimed to assess the scientific evidence for the use of Hwanglyeonhaedok-tang, a traditional herbal formula, in the treatment of rhinitis, and prepare the basis for the investigational new drug application by analyzing the experimental studies. Methods : Ten electronic databases were searched up to December, 2016 without language limitation. Experimental studies on the anti-inflammatory and anti-allergic effects of Hwanglyeonhaedok-tang against rhinitis were included. We extracted data about study design, characteristics of intervention, outcomes, and pharmacological effects from the included studies and summarized them. Results : Eight hundred and thirty-three potentially relevant studies were identified, of which 18 experimental studies met our inclusion criteria. Of 18 included studies, 5 had conducted cell viability test, and all studies had reported that Hwanglyeonhaedok-tang was non-cytotoxic. Hwanglyeonhaedok-tang exhibits anti-inflammatory effect by regulating the inflammation-related cytokines including nitric oxide(NO), prostaglandin $E_2(PGE_2)$, interleukin-6(IL-6), and tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ in vitro and in vivo. Hwanglyeonhaedok-tang exhibits anti-allergic effect by suppressing eosinophil, and histamine levels. Hwanglyeonhaedok-tang helps in the recovery of nasal mucous membrane by supressing goblet cells, heat shock protein 70, and substance P. Conclusions : This study suggests that Hwanglyeonhaedok-tang has the potential to be developed as therapeutic agent for rhinitis. Further experimental and clinical studies needed to be performed to prove the safety and efficacy.

Screening Differential Expressions of Defense-related Responses in Cold-treated 'Kyoho' and 'Campbell Early' Grapevines

  • Ahn, Soon Young;Kim, Seon Ae;Han, Jae Hyun;Kim, Seung Heui;Yun, Hae Keun
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.275-281
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    • 2013
  • Low temperature is one of the major environmental factors that affect productivity including reduced growth and budding of vines, and changes of metabolic processes in grape (Vitis spp.). To screen the specific expression of abiotic stress-related genes against cold treatment in 'Kyoho' and 'Campbell Early' grapevines, expression of various defense-related genes was investigated by RT-PCR and real-time PCR. Among the 67 genes analyzed by RT-PCR and real-time PCR, 17 and 16 types of cDNA were up-regulated, while 5 and 6 types were down-regulated in cold-treated 'Kyoho' and 'Campbell Early' grapevines, respectively. Genes encoding carotene (Cart3564 and Cart4472), chalcone isomerase (CHI), cytochrome P450 (CYP), flavonol synthase (FLS), endo-${\beta}$-glucanase precursor (Glu), glutathione peroxidase (GPX), glutathione-S-transferase (GST), leucine-rich repeats (LRR), manganese superoxide dismutase (Mn-SOD), phenylalanine ammonia lyase (PAL), polygalacturonase-inhibiting protein (PGIP), proline rich protein 2 (PRP2), small heat shock protein (sHSP), temperature induced lipocalin (TIL), and thaumatin-like protein (TLP) were up-regulated, while those encoding CBF like transcription factor (CBF1), chitinase-like protein (CLP), cold induced protein (CIP), glycerol-3-phosphate acyltransferase (GPAT), and mitogen-activated protein kinase (MAPK) were down-regulated by low temperature treatment in both in 'Kyoho' and 'Campbell Early'.

Molecular and Genomic Approaches on Nickel Toxicity and Carcinogenicity

  • Seo, Young-Rok;Kim, Byung-Joo;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.1 no.2
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    • pp.73-77
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    • 2005
  • Nickel is the one of potent environmental, the occupational pollutants and the classified human carcinogens. It is a serious hazard to human health, when the metal exposure. To prevent human diseases from the heavy metals, it is seemingly important that understanding of how nickel exerts their toxicity and carcinogenic effect at a molecular and a genomic level. The process of nickel absorption has been demonstrated as phagocytosis, iron channel and diffusion. Uptaked nickel has been suggested to induce carcinogenesis via two pathways, a direct DNA damaging pathway and an indirect DNA damaging pathway. The former was originated from the ability of metal to generate Reactive Oxygen Species (ROS) and the reactive intermediates to interact with DNA directly. Ni-generated ROS or Nickel itself, interacts with DNAs and histones to cause DNA damage and chromosomal abnormality. The latter was originated from an indirect DNA damage via inhibition of DNA repair, or condensation and methylation of DNA. Cells have ability to protect from the genotoxic stresses by changing gene expression. Microarray analysis of the cells treated with nickel or nickel compounds, show the specific altered gene expression profile. For example, HIF-I (Hypoxia-Inducible Factor I) and p53 were well known as transcription factors, which are upregulated in response to stress and activated by both soluble and insoluble nickel compounds. The induction of these important transcription factors exert potent selective pressure and leading to cell transformation. Genes of metallothionein and family of heat shock proteins which have been known to play role in protection and damage control, were also induced by nickel treatment. These gene expressions may give us a clue to understand of the carcinogenesis mechanism of nickel. Further discussions on molecular and genomic, are need in order to understand the specific mechanism of nickel toxicity and carcinogenicity.

The Expression of Adipophilin Is Frequently Found in Solid Subtype Adenocarcinoma and Is Associated with Adverse Outcomes in Lung Adenocarcinoma

  • Shin, Sun Ah;Na, Hee Young;Choe, Ji Young;Chung, Doohyun;Park, Mira;Oh, Sohee;Kim, Ji Eun
    • Journal of Pathology and Translational Medicine
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    • v.52 no.6
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    • pp.357-362
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    • 2018
  • Background: The up-regulation of the lipogenic pathway has been reported in many types of malignant tumors. However, its pathogenic role or clinical significance is not fully understood. The objective of this study was to examine the expression levels of adipophilin and related hypoxic signaling proteins and to determine their prognostic impacts and associations with the pathologic characteristics of lung adenocarcinoma. Methods: Expression levels of adipophilin, heat shock protein 27 (HSP27), carbonic anhydrase IX, and hypoxia-inducible factor $1{\alpha}$ were examined by immunohistochemical staining using tissue microarray blocks. Correlations between protein expression levels and various clinicopathologic features were analyzed. Results: A total of 230 cases of primary adenocarcinoma of the lung were enrolled in this study. Adipophilin expression was more frequent in males and with the solid histologic type. It was correlated with HSP27 expression. Patients with adipophilin-positive adenocarcinoma showed a shorter progression-free survival (PFS) (median PFS, 17.2 months vs 18.4 months) in a univariable survival analysis, whereas HSP27 positivity correlated with favorable overall survival (OS) and PFS. In a multivariable analysis, adipophilin and HSP27 were independent prognostic markers of both OS and PFS. Conclusions: Activated lipid metabolism and the hypoxic signaling pathway might play a major role in the progression of lung adenocarcinoma, especially in the solid histologic type.