Analysis of RNA for transcripts for catalase and SP71 in rat hearts after in vivo hyperthermia

1991 ◽  
Vol 69 (5-6) ◽  
pp. 375-382 ◽  
Author(s):  
R. William Currie ◽  
Robert M. Tanguay
Keyword(s):  
Heat Shock ◽  
Heat Shock Response ◽  
Catalase Activity ◽  
Mrna Level ◽  
Regulation Of Transcription ◽  
Rat Tissues ◽  
Mrna Levels ◽  
Control Heart ◽  
Shock Response ◽  
Human Hsp70

Hyperthermic stress induces synthesis of the major inducible (heat) stress protein (SP71) in all rat tissues. In addition, there is an increase in catalase activity in hearts at 24 and 48 h after the induction of the heat shock response. To more precisely define some of the molecular aspects of the induction of the heat shock response in hearts, we examined mRNA levels for the catalase, SP71 and HSP27. RNA was isolated from control hearts and at various time periods (0–24 h) of recovery after brief hyperthermic treatment and was analyzed by Northern blot analysis using as probes cDNA sequences for rat liver catalase, human HSP70 (inducible), and human HSP27. There was no detectable change in mRNA for catalase after heat shock or during recovery. Hyperthermic stress has no apparent effect on the regulation of transcription of mRNA coding for catalase, indicating that the increase in catalase activity is either translationally or post-translationally regulated. The human HSP70 cDNA did not hybridize to control heart RNA, but did hybridize to SP71 transcripts at 0, 1.5, and 3 h post heat shock. The mRNA level for SP71 peaked at 1.5 h, was reduced at 3 h, and became almost undetectable at 6 h post heat shock. Similarly, the human HSP27 cDNA did not hybridize to control heart RNA, but did hybridize to transcripts for HSP27 at 0, 1.5, 3, and up to 15 h post heat shock. Maximal signal for HSP27 was at 3 h post heat shock and was sharply reduced at 6 h post heat shock. The accumulation of transcripts for SP71 and HSP27 after hyperthermic stress is rapid, and degradation of the transcripts is almost complete by 6 h post heat shock.Key words: heat shock proteins, hyperthermia, catalase, heart, RNA.

Induction of the heat shock response in Arabidopsis by heat shock protein 70 inhibitor VER-155008

2019 ◽  
Vol 46 (10) ◽  
pp. 925
Author(s):  
Erina Matsuoka ◽  
Naoki Kato ◽  
Masakazu Hara
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Heat Shock Response ◽  
Wheat Germ ◽  
Heat Shock Protein 90 ◽  
Hsp90 Inhibitor ◽  
Wheat Germ Extract ◽  
Chlorophyll Contents ◽  
Shock Response ◽  
Human Hsp70

The heat shock protein 90 (HSP90) inhibitor, geldanamycin, is a chemical inducer of the heat shock response (HSR) in Arabidopsis. Geldanamycin is thought to activate the heat shock signal by dissociating the HSP90-heat shock factor (HSF) complex. Recent studies have indicated that plant HSP70 is also associated with HSF, suggesting that inhibition of HSP70 may induce the HSR. However, no studies have been conducted to test this hypothesis. Here, we found that a specific HSP70 inhibitor VER-155008 activated the promoter of a small HSP gene (At1 g53540, HSP17.6C-CI) of Arabidopsis, which was shown to be activated by geldanamycin and other HSP90 inhibitors. The production of HSP17.6C-CI, HSP70 and HSP90.1 proteins in Arabidopsis was enhanced by the addition of VER-155008. The reduction of chlorophyll contents by heat shock was ameliorated by VER-155008. Chaperone analyses indicated that VER-155008 inhibited the chaperone activities of wheat germ extract and human HSP70/HSP40, respectively. These results suggest that the inhibition of HSP70 by VER-155008 enhanced the heat tolerance of Arabidopsis by inducing the HSR in the plant.

scholarly journals The heat shock response in HeLa cells is accompanied by elevated expression of the c-fos proto-oncogene.

1987 ◽  
Vol 7 (10) ◽  
pp. 3452-3458 ◽  
Author(s):  
G K Andrews ◽  
M A Harding ◽  
J P Calvet ◽  
E D Adamson
Keyword(s):  
Heavy Metals ◽  
Protein Synthesis ◽  
Heat Stress ◽  
Heat Shock ◽  
Hela Cells ◽  
Heat Shock Response ◽  
Mrna Level ◽  
Hsp 70 ◽  
Shock Response ◽  
Elevated Expression

Several known inducers of the heat shock response (heat stress, arsenite, and heavy metals) were shown to cause a significant elevation of c-fos mRNA in HeLa cells. Heat stress resulted in a time- and temperature-dependent prolonged elevation in the level of c-fos mRNA, which was accompanied by increased translation of c-fos protein and its appearance in the nucleus. Elevated expression of c-fos during heat stress was paralleled by induction of hsp 70 mRNA, while levels of c-myc and metallothionein mRNAs declined. Treatment of HeLa cells with arsenite or heavy metals also resulted in increased levels of hsp 70, as well as c-fos mRNA. Although elevated expression of c-fos was prevented by inhibitors of RNA synthesis, analysis of relative rates of gene transcription showed that during heat stress there was a negligible change in c-fos transcription. Therefore, the enhanced expression of c-fos during the heat shock response is likely to occur primarily through posttranscriptional processes. Cycloheximide was also shown to significantly increase the c-fos mRNA level in HeLa cells. There results are consistent with the observation that these inducers of the heat shock response, as well as cycloheximide, repress protein synthesis and suggest that the increase in the level of c-fos mRNA is caused by an inhibition of protein synthesis. This supports the hypothesis that c-fos mRNA is preferentially stabilized under conditions which induce the heat shock response, perhaps by decreased synthesis of a short-lived protein which regulates c-fos mRNA turnover.

scholarly journals The effect of aging on the autophagic and heat shock response in human peripheral blood mononuclear cells

2018 ◽  
Vol 105 (3) ◽  
pp. 247-256 ◽  
Author(s):  
JJ McCormick ◽  
TA VanDusseldorp ◽  
CG Ulrich ◽  
RL Lanphere ◽  
K Dokladny ◽  
...  
Keyword(s):  
Heat Shock ◽  
Protein Expression ◽  
Peripheral Blood Mononuclear Cells ◽  
Heat Shock Response ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Mrna Levels ◽  
Peripheral Blood Mononuclear ◽  
Shock Response ◽  
Blood Mononuclear Cells

Autophagy is a lysosome degradation pathway through which damaged organelles and macromolecules are degraded within the cell. A decrease in activity of the autophagic process has been linked to several age-associated pathologies, including triglyceride accumulation, mitochondrial dysfunction, muscle degeneration, and cardiac malfunction. Here, we examined the differences in the autophagic response using autophagy-inducer rapamycin (Rapa) in peripheral blood mononuclear cells (PBMCs) from young (21.8 ± 1.9 years) and old (64.0 ± 3.7 years) individuals. Furthermore, we tested the interplay between the heat shock response and autophagy systems. Our results showed a significant increase in LC3-II protein expression in response to Rapa treatment in young but not in old individuals. This was associated with a decreased response in MAP1LC3B mRNA levels, but not SQSTM1/p62. Furthermore, HSPA1A mRNA was upregulated only in young individuals, despite no differences in HSP70 protein expression. The combined findings suggest a suppressed autophagic response following Rapa treatment in older individuals.

scholarly journals Urocortin 3 overexpression reduces ER stress and heat shock response in 3T3-L1 adipocytes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sina Kavalakatt ◽  
Abdelkrim Khadir ◽  
Dhanya Madhu ◽  
Heikki A. Koistinen ◽  
Fahd Al-Mulla ◽  
...  
Keyword(s):  
Stress Response ◽  
Heat Shock ◽  
Er Stress ◽  
Glucose Uptake ◽  
Heat Shock Response ◽  
Cellular Stress Response ◽  
Mrna Levels ◽  
Obesity And Diabetes ◽  
Shock Response ◽  
Urocortin 3

AbstractThe neuropeptide urocortin 3 (UCN3) has a beneficial effect on metabolic disorders, such as obesity, diabetes, and cardiovascular disease. It has been reported that UCN3 regulates insulin secretion and is dysregulated with increasing severity of obesity and diabetes. However, its function in the adipose tissue is unclear. We investigated the overexpression of UCN3 in 3T3-L1 preadipocytes and differentiated adipocytes and its effects on heat shock response, ER stress, inflammatory markers, and glucose uptake in the presence of stress-inducing concentrations of palmitic acid (PA). UCN3 overexpression significantly downregulated heat shock proteins (HSP60, HSP72 and HSP90) and ER stress response markers (GRP78, PERK, ATF6, and IRE1α) and attenuated inflammation (TNFα) and apoptosis (CHOP). Moreover, enhanced glucose uptake was observed in both preadipocytes and mature adipocytes, which is associated with upregulated phosphorylation of AKT and ERK but reduced p-JNK. Moderate effects of UCN3 overexpression were also observed in the presence of 400 μM of PA, and macrophage conditioned medium dramatically decreased the UCN3 mRNA levels in differentiated 3T3-L1 cells. In conclusion, the beneficial effects of UCN3 in adipocytes are reflected, at least partially, by the improvement in cellular stress response and glucose uptake and attenuation of inflammation and apoptosis.

Acquisition and decay of heat-shock-enhanced postischemic ventricular recovery

1990 ◽  
Vol 259 (2) ◽  
pp. H424-H431 ◽  
Author(s):  
M. Karmazyn ◽  
K. Mailer ◽  
R. W. Currie
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Heat Shock Response ◽  
Strong Relationship ◽  
Male Rats ◽  
Rat Tissues ◽  
Ischemia And Reperfusion ◽  
Isolated Hearts ◽  
Shock Response ◽  
Postischemic Recovery

Hyperthermia induces the synthesis of the 71-kDa heat-shock protein (heat-shock response) in all rat tissues, including heart. We examined whether induction of the heat-shock response alters the response of isolated hearts to ischemia and reperfusion. Anesthetized male rats were pretreated with 15 min of hyperthermia (42 degrees C) and then recovered for 0, 24, 48, 96, or 192 h. Hearts were isolated from control and hyperthermia-treated rats and retrogradely perfused. Greatest recovery occurred in 48-h postheat-shock hearts; after 30 min of reperfusion there was a 38, 62, and 62% recovery of force, +dF/dt, and -dF/dt, respectively, and 17, 36, and 30% recovery, respectively, for the control hearts. Creatine kinase efflux during reperfusion was reduced by 75% for 24-h postheat-shock hearts. The antioxidative enzyme catalase was increased 24, 48, and 96 h posthyperthermia. Treatment of rats with 3-amino-1,2,4-triazole (1 g/kg body wt), which irreversibly inactivates catalase, 30 min before isolation of hearts, abolished the hyperthermia-induced enhancement of postischemic recovery. These results show a strong relationship between the acquisition and decay of the enhanced postischemic ventricular recovery and the hyperthermic induction of the heat-shock response indicated by the accumulation of heat-shock protein HSP71 (mol mass 71 kDa) and the increase in catalase activity.

Heat shock protects L6 myotubes from catabolic effects of dexamethasone and prevents downregulation of NF-κB

2001 ◽  
Vol 281 (4) ◽  
pp. R1193-R1200 ◽  
Author(s):  
Guangju Luo ◽  
Xiaoyan Sun ◽  
Eric Hungness ◽  
Per-Olof Hasselgren
Keyword(s):  
Heat Shock ◽  
Protein Degradation ◽  
Heat Shock Response ◽  
Muscle Cells ◽  
Binding Activity ◽  
Mrna Levels ◽  
Dna Binding Activity ◽  
Heat Shock Stress ◽  
L6 Myotubes ◽  
Shock Response

Glucocorticoids are the most important mediator of muscle cachexia in various catabolic conditions. Recent studies suggest that the transcription factor NF-κB acts as a suppressor of genes in the ubiquitin-proteasome proteolytic pathway and that glucocorticoids increase muscle proteolysis by downregulating NF-κB activity. The heat shock (stress) response, characterized by the induction of heat shock proteins, confers a protective effect against a variety of harmful stimuli. In the present study, we tested the hypothesis that the heat shock response protects muscle cells from the catabolic effects of dexamethasone and prevents downregulation of NF-κB. Cultured L6 myotubes were subjected to heat shock (43°C for 1 h) followed by recovery at 37°C for 1 h. Thereafter, cells were treated for 6 h with 1 μM dexamethasone, during which period protein degradation was measured as release of TCA-soluble radioactivity from proteins that had been prelabeled with [3H]tyrosine. Heat shock resulted in increased protein and mRNA levels for heat shock protein 70. The increase in protein degradation induced by dexamethasone was prevented in cells expressing the heat shock response. In the same cells, dexamethasone-induced downregulation of NF-κB DNA binding activity was blocked. The present results suggest that the heat shock response may protect muscle cells from the catabolic effects of dexamethasone and that this effect of heat shock may be related to inhibited downregulation of NF-κB activity.

The heat shock response in HeLa cells is accompanied by elevated expression of the c-fos proto-oncogene

1987 ◽  
Vol 7 (10) ◽  
pp. 3452-3458
Author(s):  
G K Andrews ◽  
M A Harding ◽  
J P Calvet ◽  
E D Adamson
Keyword(s):  
Heavy Metals ◽  
Protein Synthesis ◽  
Heat Stress ◽  
Heat Shock ◽  
Hela Cells ◽  
Heat Shock Response ◽  
Mrna Level ◽  
Hsp 70 ◽  
Shock Response ◽  
Elevated Expression

Several known inducers of the heat shock response (heat stress, arsenite, and heavy metals) were shown to cause a significant elevation of c-fos mRNA in HeLa cells. Heat stress resulted in a time- and temperature-dependent prolonged elevation in the level of c-fos mRNA, which was accompanied by increased translation of c-fos protein and its appearance in the nucleus. Elevated expression of c-fos during heat stress was paralleled by induction of hsp 70 mRNA, while levels of c-myc and metallothionein mRNAs declined. Treatment of HeLa cells with arsenite or heavy metals also resulted in increased levels of hsp 70, as well as c-fos mRNA. Although elevated expression of c-fos was prevented by inhibitors of RNA synthesis, analysis of relative rates of gene transcription showed that during heat stress there was a negligible change in c-fos transcription. Therefore, the enhanced expression of c-fos during the heat shock response is likely to occur primarily through posttranscriptional processes. Cycloheximide was also shown to significantly increase the c-fos mRNA level in HeLa cells. There results are consistent with the observation that these inducers of the heat shock response, as well as cycloheximide, repress protein synthesis and suggest that the increase in the level of c-fos mRNA is caused by an inhibition of protein synthesis. This supports the hypothesis that c-fos mRNA is preferentially stabilized under conditions which induce the heat shock response, perhaps by decreased synthesis of a short-lived protein which regulates c-fos mRNA turnover.

Abstract 106: A Novel Small Molecule Protects Against Myocardial Ischemia and Doxorubicin-Induced Cardiomyopathy Through Heat Shock Response and Proteostasis Regulation In Vivo

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Sudhish Sharma ◽  
Savitha Deshmukh ◽  
Manuela Zampino ◽  
Brandon Walker ◽  
Sunjay Kaushal
Keyword(s):  
Heat Shock ◽  
Small Molecule ◽  
Heat Shock Response ◽  
Myocardial Injury ◽  
Heart Function ◽  
Myocardial Infarct Size ◽  
Mrna Levels ◽  
Cell Based Therapy ◽  
Shock Response

Background: A potentially new non-cell-based therapy is the systemic delivery of small molecules to revive or protect injured myocardium. By screening a drug library, we identified a novel small molecule, sk421, and tested its effectiveness in two myocardial injury models in vivo: a myocardial infarction model and doxorubicin (DOX)-induced cardiomyopathy model. Results: Sk421 significantly increased cellular viability of DOX-injured cardiomyocytes by preventing apoptosis in vitro, demonstrated by assays for MTT (p=0.042), annexin-V, and active caspase-3 (p=0.004). Sk421 increased the concentrations of the GATA4 and the heat shock proteins (HSPs) in cardiomyocytes (by 1.5-fold), and fibroblasts (by 1.2-fold), but was unable to rescue DOX-injured fibroblast cells depleted of HSPs. Sk421 maintained the total proteosome by a 3.5-fold inhibition of ubiquitinized GATA4 degradation, as shown by immunoprecipitation assays. Oral administration of sk421(4mg/kg) rescued DOX-injured mice by significantly preserving cardiac function, weight gain, cardiomyocyte lesion size and abundance, and by reducing overall mortality (20% in treated groups) compared to control DOX-injured mice (80% in untreated groups, p<0.03). The levels of GATA4 and HSPs were increased in the sk421-rescued mice. In the MI model, (n=5/group), sk421 reduced myocardial infarct size, and protected heart function evaluated by ejection fraction 45.8 ± 2.4 vs. 34.8 ± 3.3, p=0.005 (day7), and 44.3 ± 2.8 vs. 34.4 ± 4.1, p=0.011 (day28). Sk421 treatment of MI rats resulted in a higher fraction of viable tissue (within the LV (70.20 ± 0.5024, n=3) as compared to only MI group (56.38 ± 3.879%, n=3) and the infiltration of CD68+ cells was found to be more in the infarct region of LV of MI group rats as compared to MI+CEL. Quantitative analysis showed a significantly higher proportion of TUNEL positive cells and mRNA levels of IL-1β, IL-6, TNF-α and MMP2 in the myocardium of MI rats compared to MI+SK421 rats. Conclusion: This study demonstrates the broad therapeutic benefit of sk421 in animal model of myocardial injury by mechanism of inducing the heat shock response and preserving GATA levels. The ease of oral delivery makes sk421 a potentially clinically useful adjunct for myocardial preservation.

scholarly journals Interfering with Different Steps of Protein SynthesisExplored by Transcriptional Profiling of Escherichia coliK-12

2003 ◽  
Vol 185 (20) ◽  
pp. 6158-6170 ◽  
Author(s):  
Jeffrey Sabina ◽  
Nir Dover ◽  
Lori J. Templeton ◽  
Dana R. Smulski ◽  
Dieter Söll ◽  
...  
Keyword(s):  
Heat Shock ◽  
Ribosomal Protein ◽  
Heat Shock Response ◽  
Ribosomal Subunit ◽  
Trna Synthetase ◽  
Transcriptional Level ◽  
Transferase Activity ◽  
Synthetase Activity ◽  
Mrna Levels ◽  
Shock Response

ABSTRACT Escherichia coli responses to four inhibitors that interfere with translation were monitored at the transcriptional level. A DNA microarray method provided a comprehensive view of changes in mRNA levels after exposure to these agents. Real-time reverse transcriptase PCRanalysis served to verify observations made with microarrays, and a chromosomal grpE::lux operon fusion was employed to specifically monitor the heat shock response. 4-Azaleucine, a competitive inhibitor of leucyl-tRNA synthetase, surprisingly triggered the heat shock response. Administration of mupirocin, an inhibitor of isoleucyl-tRNA synthetase activity, resulted in changes reminiscent of the stringent response. Treatment with kasugamycin and puromycin (targeting ribosomal subunit association as well as its peptidyl-transferase activity) caused accumulation of mRNAs from ribosomal protein operons. Abundant biosynthetic transcripts were often significantly diminished after treatment with any of these agents. Exposure of a relA strain to mupirocin resulted in accumulation of ribosomal protein operon transcripts. However, the relA strain's response to the other inhibitors was quite similar to that of the wild-type strain.

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