scholarly journals Involvement of HSP Synthesis and Protease Inhibitors in Heat Shock-induced Cucumber Seedling Chilling Tolerance

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 775F-775
Author(s):  
Paul H. Jennings ◽  
Ann Fitzpatrick
Keyword(s):  
Protein Synthesis ◽  
Heat Shock ◽  
Protease Inhibitors ◽  
Actinomycin D ◽  
Chilling Tolerance ◽  
Cucumber Seedling ◽  
Shock Effect ◽  
Seedling Roots ◽  
Germinating Seeds ◽  
Heat Shock Induction

Heat shock induction of chilling tolerance in cucumber seedlings is not blocked by inhibitors of protein synthesis. Treatment of germinating seeds with cycloheximide and actinomycin-D, prior to heat shock and chilling, does not block the heat shock induction of chilling tolerance, while the inhibitors alone promote chilling tolerance of seedling roots. To test whether the heat shock effect might be acting on proteases, two protease inhibitors (bestatin and PMSF) were tested for their ability to induce chilling tolerance. Although PMSF slowed germination, it still provided protection against chilling, but bestatin was much more effective.

scholarly journals The Effect of Heat Shock on the Induction of Chilling Tolerance in Cucumber Seedling Roots

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 851F-851 ◽  
Author(s):  
Hua Zhang ◽  
Paul H. Jennings
Keyword(s):  
Molecular Weight ◽  
Heat Shock ◽  
Protein Profile ◽  
Chilling Tolerance ◽  
Low Molecular Weight ◽  
Cucumber Seedling ◽  
Sds Page ◽  
Seedling Roots ◽  
Shock Proteins ◽  
Heat Shock Induction

Heat shock was applied to 32-h-old cucumber seedlings before chilling at 2.5C. Two cultivars, `Poinsett 76' and `Ashley', with different chilling tolerances, were tested. Using root growth after chilling as a measure of chilling tolerance, three heat shock regimes were found to induce chilling tolerance in both cultivars, with the most effective and uniform induction by heat shock at 40C for 3 h. `Ashley', the more chilling tolerant cultivar, exhibited a greater response to heat shock induction of chilling tolerance than `Poinsett 76'. Protein samples from roots were subjected to SDS-PAGE. Three low molecular weight heat shock proteins accumulated to a greater extent in the protein profile of heat-shocked `Ashley' roots. No such increase was found in the `Poinsett 76' roots. The induction of low molecular weight HSPs are discussed in relation to the heat-shock induction of chilling tolerance.

scholarly journals The Effect of Heat Shock, ABA, and Salicylic Acid on Induction of Chilling Tolerance in Cucumber Seedling Roots

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 913H-914
Author(s):  
Meng-Yee Tee ◽  
Paul H. Jennings
Keyword(s):  
Salicylic Acid ◽  
Heat Shock ◽  
Chilling Injury ◽  
Chilling Tolerance ◽  
Cucumber Seedling ◽  
Crop Species ◽  
Growth Effects ◽  
Seedling Roots ◽  
Treatment Mechanisms ◽  
Germinated Seeds

Chilling injury can be a serious problem during field germination of sensitive crop species. Because heat shock has been shown to induce chilling tolerance of germinating cucumber seeds, an experiment was initiated to determine the effectiveness of other treatments. Cucumber seeds germinated 20 to 24 h were either heat-shocked at 50C for 2 min or treated with ABA or salicylic acid for 4 h. Following treatment, the germinated seeds were chilled at 2C for 96, 120, or 144 h and then incubated at 25C to determine growth effects on the developing root. All treatments induced chilling tolerance compared to the controls, with ABA and heat shock being most effective after chilling. There did not appear to be an additive response when heat shock was used in combination with ABA. The evidence for different treatment mechanisms will be discussed.

scholarly journals Effects of Chilling, Ethanol, and Heat Shock on Enzyme Expression in Cucumber Seedling Roots

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 912D-912
Author(s):  
Windy A. Boyd ◽  
Paul H. Jennings
Keyword(s):  
Heat Shock ◽  
Catalase Activity ◽  
Chilling Tolerance ◽  
Activity Levels ◽  
Cucumber Seedling ◽  
Enzyme Expression ◽  
Cucumber Seedlings ◽  
Before And After ◽  
Seedling Roots ◽  
Chilling Period

Chilling-sensitive cucumber seedlings that are treated with ethanol or heat-shocked have shown an increase in chilling tolerance. The mechanisms that regulate this response have not been identified. Cucumber seeds were germinated for 24 h and then treated with 500 mM ethanol for 2 h or heat-shocked at 40C for 3 h. Immediately after treatment, roots were excised and catalase activity was assayed. Another set of control and treated seeds were chilled for 72 h and catalase was assayed at the end of the chilling period. Comparisons will be presented between catalase activity levels before and after chilling, as well as between the control and treated groups. Treatments of ethanol and heat shock resulted in an increase in catalase activity when compared to controls.

Expression of a Drosophila heat shock protein in mammalian cells: transient association with nucleoli after heat shock

1984 ◽  
Vol 307 (1132) ◽  
pp. 301-307 ◽  
Keyword(s):  
Protein Synthesis ◽  
Heat Shock ◽  
Mammalian Cells ◽  
Actinomycin D ◽  
Future Studies ◽  
Cos Cells ◽  
Single Protein ◽  
Cellular Components ◽  
Multiple Interactions

We transformed mouse L cells with a cloned Drosophila hsp70 gene and obtained cells with either heat-inducible or constitutively expressed copies of the gene. The distribution of hsp70 in these cells was examined by indirect immunofluorescence with monoclonal antibodies specific to the Drosophila protein. In constitutive cells, hsp70 was present in both cytoplasm and nucleus. After heat shock, the nuclear hsp70 was transiently concentrated in nucleoli, from which it had previously been excluded; the cytoplasmic hsp70 moved to a perinuclear location, a result consistent with it being associated with intermediate filaments of the cytoskeleton. The nucleolar migration took several hours and was partly inhibited by actinomycin D, but was independent of protein synthesis; it may reflect binding to newly-synthesized rRNA. Drosophila hsp70 was also expressed from replicating plasmids in monkey COS cells, and was found to be concentrated in nuclei even at low temperature. Migration to nucleoli occurred after heat shock. These results indicate that a single protein can have multiple interactions with cellular components, and form the basis for future studies of these interactions by in vitro mutagenesis and expression of the hsp70 gene.

scholarly journals Duration and Persistence of Heat Shock Induction of Chilling Tolerance in Cucumber Seedling Roots

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 538E-538
Author(s):  
Hua Zhang ◽  
Paul H. Jennings
Keyword(s):  
Heat Shock ◽  
Root Growth ◽  
Electrolyte Leakage ◽  
Chilling Tolerance ◽  
Shock Duration ◽  
Dependent Manner ◽  
Heat Shocks ◽  
Shock Proteins ◽  
Membrane Peroxidation ◽  
Heat Shock Induction

The effects of heat shock duration and persistence on the induction of chilling tolerance in cucumber roots were studied using total root growth, electrolyte leakage, and membrane peroxidation as injury indices after chilling. Heat shock reduced the chilling induced electrolyte leakage, decreased membrane peroxidation as measured by MDA content, and resulted in a greater total root growth after chilling compared to the control. Heat shocks at 40°C, applied to 36 hr germinated seedlings for time periods from 1 to 15 hr, all resulted in an increase in chilling tolerance in a time-dependent manner. The heat shock induction of chilling tolerance is most effective when heat shock was imposed immediately before chilling, but the effect is persistent even 32 hr after heat shock when seedlings are held at 25°C before chilling. The possible mechanism of heat shock effect and its persistence will be discussed in relation to heat shock proteins and antioxidant enzyme systems.

Recovery of normal protein synthesis in heat-shocked chicken myotubes by liposome-mediated transfer of mRNAs

1985 ◽  
Vol 63 (3) ◽  
pp. 231-235 ◽  
Author(s):  
Jnanankur Bag
Keyword(s):  
Protein Synthesis ◽  
Heat Shock ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
De Novo ◽  
Critical Role ◽  
Recovery Process ◽  
De Novo Synthesis ◽  
Normal Pattern ◽  
Large Unilamellar Vesicles

Exposure of chicken myotube culture to 45 °C induced the synthesis of three heat-shock polypeptides of 25 000, 65 000, and 81 000 daltons. Recovery to the normal pattern of protein synthesis was judged by the decrease in the synthesis of heat-shock polypeptides. This recovery to normal protein synthesis required de novo synthesis of mRNAs for normal cellular proteins. Inhibition of RNA synthesis by actinomycin D during recovery at 37 °C blocked the recovery process and resulted in the continued synthesis of heat-shock polypeptides. Large unilamellar vesicles were used to examine the effect of delivery of mRNAs isolated from both normal and heat-shocked myotubes on the recovery of these cells from heat-shock treatment. The results presented here show that liposome-mediated delivery of normal mRNAs to heat-shocked cells relieved the block of recovery by actinomycin. On the other hand, when mRNAs from heat-shocked cells were used during recovery, the synthesis of heat-shock polypeptides was stimulated. These observations suggest that the relative abundance of mRNAs in the cytoplasm plays a critical role in regulating protein synthesis in chicken myotube cultures.

EFFECT OF ACTINOMYCIN D ON TSH-INDUCED STIMULATION OF THYROIDAL PROTEIN SYNTHESIS IN THE RAT

1974 ◽  
Vol 77 (1) ◽  
pp. 64-70 ◽  
Author(s):  
Gustav Wägar
Keyword(s):  
Protein Synthesis ◽  
Actinomycin D ◽  
The Other ◽  
Leucine Incorporation ◽  
Short Term ◽  
Other Hand ◽  
Thyroid Glands ◽  
Translational Level ◽  
Stimulation Of

ABSTRACT Whether the short-term regulation of thyroidal protein synthesis by TSH occurs at the transcriptional or the translational level was tested by measuring the effect of actinomycin D (act D) on the TSH-induced stimulation of L-14C-leucine incorporation into the thyroidal proteins of rats. TSH was injected 6 h before the rats were killed. The thyroid glands were then removed and incubated in vitro in the presence of L-14C-leucine for 2 h. The pronounced stimulation of leucine incorporation in the TSH-treated animals was depressed as compared with controls but still significant even when the animals had been pre-treated with 100 μg act D 24 and 7 h before sacrifice. On the other hand, act D strongly decreased incorporation of 3H-uridine into RNA. Short-term regulation of thyroidal protein synthesis by TSH appears to be partly but not wholly dependent on neosynthesis of RNA. Hence regulation may partly occur at the translation level of protein synthesis.

Dimethyl-sulfoxide is a Suitable Solvent for Fluorescent Microscopy Detection of Medium and Strong Heat Shock Inductors Using Transgenic Zebrafish

2018 ◽  
Vol 69 (2) ◽  
pp. 337-340
Author(s):  
Vlad Preluca ◽  
Bogdan Horatiu Serb ◽  
Sanda Marchian ◽  
Diter Atasie ◽  
Mihaela Cernusca Mitariu ◽  
...  
Keyword(s):  
Heat Shock ◽  
Dimethyl Sulfoxide ◽  
Heat Shock Response ◽  
Protein Misfolding ◽  
Fluorescent Microscopy ◽  
Screening Tests ◽  
Transgenic Zebrafish ◽  
Fluorescent Intensity ◽  
Shock Response ◽  
Heat Shock Induction

Heat shock inductors have potential as treatment for degenerative and protein misfolding diseases. Dimethyl-sulfoxide is widely used as a solvent in pharmacological screening tests and has been shown to have heat shock induction effects. Transgenic Tg (hsp70l:EGFP-HRAS_G12V)io3(AB) zebrafish larvae were exposed for 24 hours to dimethyl-sulfoxide in concentratios of 0.1-2%, and to moderate heat shock inductors pentoxifylline and tacrolimus. Positive controls were exposed to 35, 38 and 40�C for 20 min, and incubated for 24 h at 28�C. Heat shock response was measured by fluorescence microscopy and signal intensity quantification in FIJI. Dimethyl-sulfoxide caused a dose-dependant increase in fluorescent intensity, but significantly lower compared with exposure to 38 and 40�C. Pentoxifylline and tacrolimus induced a significantly higher increase in fluorescence compared with 0.5% dimethyl-sulfoxide. Thus, although dimethyl-sulfoxide has independent heat shock induction effects, concentrations of up to 0.5% are suitable for heat shock response screening tests.

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