scholarly journals Gene expression changes after heat shock of procyclic-form Trypanosoma brucei suggest that stress has a role in differentiation to mammalian-infective forms

2016 ◽  
Author(s):  
Igor Minia ◽  
Clementine Merce ◽  
Monica Terrao ◽  
Christine Clayton
Keyword(s):  
Heat Shock ◽  
Salivary Glands ◽  
Zinc Finger Protein ◽  
Diurnal Variations ◽  
Tsetse Flies ◽  
Granule Formation ◽  
Temperature Variations ◽  
Warm Blood ◽  
Heat Shocks ◽  
Polysomal Fraction

AbstractTrypanosome procyclic forms multiply in the midgut of Tsetse flies, and are routinely cultured at 27°C. Heat shocks of 37°C and above result in general inhibition of translation, and severe heat shock (41°C) results in sequestration of mRNA in granules. The mRNAs that are bound by the zinc-finger protein ZC3H11, including those encoding refolding chaperones, escape heat-induced translation inhibition.a At 27°C, ZC3H11 mRNA is predominantly present as an untranslated cytosolic messenger ribonucleoprotein particle, but after heat shocks of 37°C - 41°C, the ZC3H11 mRNA moves into the polysomal fraction. To investigate the scope and specificities of heat-shock translational regulation and granule formation, we analysed the distributions of mRNAs on polysomes at 27C and after 1 hour at 39°C, and the mRNA content of 41°C heat shocks granules. We found that that mRNAs that bind to ZC3H11 remained in polysomes at 39°C and were protected from sequestration in granules at 41°C. As previously seen for starvation stress granules, the mRNAs that encode ribosomal proteins were excluded from heat-shock granules. Seventy mRNAs moved towards the polysomal fraction after the 39°C heat shock; surprisingly, many of these are also increased when trypanosomes migrate to the Tsetse salivary glands. It therefore seems possible that in the wild, temperature changes due to diurnal variations and periodic intake of warm blood might influence the efficiency with which procyclic forms develop into mammalian-infective formsAuthor summaryWhen trypanosomes are inside tsetse flies, they have to cope with temperature variations from below 20°C up to nearly 40°C, due to diurnal variations and periodic intake of warm blood. The procyclic forms, which usually multiply in the midgut, are routinely cultured at 27°C in the laboratory. When they are heated to temperatures of 37°C and above, they shut down protein production, and at 41°C, mRNAs aggregate into granules. We show here that quite a large number of mRNAs are not included in granules and continue to be used for making proteins. Some of the proteins that continue to be made are needed in order to defend the cells against the effects of heat shock. Interestingly, however, a moderate heat shock stimulates expression of genes needed for the parasites to develop further into forms that can colonise the salivary glands. It thus seems possible that in the field, temperature variations might influence the efficiency with which of trypanosomes in tsetse flies become infective for mammals.

scholarly journals Mitochondrial responses towards intermittent heat shocks in the Eastern Oyster, Crassostrea virginica

2021 ◽  
Author(s):  
Georges Hraoui ◽  
Sophie Breton ◽  
Gilles Miron ◽  
Luc H. Boudreau ◽  
Florence Hunter-Manseau ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Heat Shock ◽  
Crassostrea Virginica ◽  
Heat Waves ◽  
Physiological Stress ◽  
Eastern Oyster ◽  
Emission Rates ◽  
Mitochondrial Functions ◽  
Heat Shocks ◽  
Single Stress

Frequent heat waves caused by climate change can cause physiological stress in many animals, particularly in sessile ectotherms such as bivalves. Most studies characterizing thermal stress in bivalves focus on evaluating the responses to a single stress event. This does not accurately reflect the reality faced by bivalves which are often subject to intermittent heat waves. Here, we investigated the effect of intermittent heat stress on mitochondrial functions of Eastern oyster Crassostrea virginica which play a key role in setting ectotherms’ thermal tolerance. Specifically, we measured changes in mitochondrial oxygen consumption and H2O2 emission rates before, during and after intermittent 7.5°C heat shocks in oysters acclimated to 15°C and 22.5°C. Our results showed that oxygen consumption was impaired following the first heat shock at both acclimation temperatures. After the second heat shock, results for oysters acclimated to 15°C indicated a return to normal. However, oysters acclimated to 22.5°C struggled more with the compounding effects of intermittent heat shocks as denoted with an increase contribution of FAD-linked substrates to mitochondrial respiration as well as high levels of H2O2 emission rates. However, both acclimated populations showed signs of potential recovery ten days after the second heat shock, reflecting a surprising resilience to heat waves by C. virginica. Thus, this study highlights the important role of acclimation in oyster's capacity to weather intermittent heat shock.

Heat shock-induced alterations in phosphorylation of the largest subunit of RNA polymerase II as revealed by monoclonal antibodies CC-3 and MPM-2

1999 ◽  
Vol 77 (4) ◽  
pp. 367-374 ◽  
Author(s):  
Sébastien B Lavoie ◽  
Alexandra L Albert ◽  
Alain Thibodeau ◽  
Michel Vincent
Keyword(s):  
Heat Shock ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Transcriptional Activity ◽  
Phosphorylation Sites ◽  
Stress Genes ◽  
Terminal Domain ◽  
Carboxy Terminal Domain ◽  
Heat Shocks ◽  
Carboxy Terminal

The phosphorylation of the carboxy-terminal domain of the largest subunit of RNA polymerase II plays an important role in the regulation of transcriptional activity and is also implicated in pre-mRNA processing. Different stresses, such as a heat shock, induce a marked alteration in the phosphorylation of this domain. The expression of stress genes by RNA polymerase II, to the detriment of other genes, could be attributable to such modifications of the phosphorylation sites. Using two phosphodependent antibodies recognizing distinct hyperphosphorylated forms of RNA polymerase II largest subunit, we studied the phosphorylation state of the subunit in different species after heat shocks of varying intensities. One of these antibodies, CC-3, preferentially recognizes the carboxy-terminal domain of the largest subunit under normal conditions, but its reactivity is diminished during stress. In contrast, the other antibody used, MPM-2, demonstrated a strong reactivity after a heat shock in most species studied. Therefore, CC-3 and MPM-2 antibodies discriminate between phosphoisomers that may be functionally different. Our results further indicate that the pattern of phosphorylation of RNA polymerase II in most species varies in response to environmental stress.Key words: RNA polymerase II, heat shock, phosphorylation, CC-3, MPM-2.

Preferential deadenylation of Hsp70 mRNA plays a key role in regulating Hsp70 expression in Drosophila melanogaster

1994 ◽  
Vol 14 (6) ◽  
pp. 3646-3659
Author(s):  
R P Dellavalle ◽  
R Petersen ◽  
S Lindquist
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Rna Processing ◽  
Hsp70 Expression ◽  
Hsp70 Mrna ◽  
Mild Heat ◽  
Endonucleolytic Cleavage ◽  
Heat Shocks ◽  
Low Efficiency ◽  
Hsp70 Synthesis

Following a standard heat shock, approximately 40% of Hsp70 transcripts in Drosophila melanogaster lack a poly(A) tail. Since heat shock disrupts other aspects of RNA processing, this observation suggested that heat might disrupt polyadenylation as well. We find, however, that as the temperature is increased a larger fraction of Hsp70 RNA is polyadenylated. Poly(A)-deficient Hsp70 RNAs arise not from a failure in polyadenylation but from the rapid and selective removal of poly(A) from previously adenylated transcripts. Poly(A) removal is highly regulated: poly(A) is (i) removed much more rapidly from Hsp70 RNAs than from Hsp23 RNAs, (ii) removed more rapidly after mild heat shocks than after severe heat shocks, and (iii) removed more rapidly after a severe heat shock if cells have first been conditioned by a mild heat treatment. Poly(A) seems to be removed by simple deadenylation rather than by endonucleolytic cleavage 5' of the adenylation site. During recovery from heat shock, deadenylation is rapidly followed by degradation. In cells maintained at high temperatures, however, the two processes are uncoupled and Hsp70 RNAs are deadenylated without being degraded. These deadenylated mRNAs are translated with low efficiency. Deadenylation therefore allows Hsp70 synthesis to be repressed even when degradation of the mRNA is blocked. Poly(A) tail shortening appears to play a key role in regulating Hsp70 expression.

scholarly journals THE RELATIONSHIP BETWEEN DEOXYRIBONUCLEIC ACID REPLICATION AND CELL DIVISION IN HEAT-SYNCHRONIZED TETRAHYMENA

1970 ◽  
Vol 46 (3) ◽  
pp. 533-543 ◽  
Author(s):  
William R. Jeffery ◽  
Kenneth D. Stuart ◽  
Joseph Frankel
Keyword(s):  
Cell Division ◽  
Heat Shock ◽  
Dna Synthesis ◽  
Deoxyribonucleic Acid ◽  
Heat Shock Treatment ◽  
Heat Shocks ◽  
Prolonged Heat ◽  
S Period ◽  
C 50 ◽  
The Relationship

The effect of supraoptimal temperature on macronuclear DNA synthesis in Tetrahymena was studied by radioautography during prolonged heat and heat-shock synchronization treatments. Prolonged heat treatments (34°C) delayed the initiation of S, but did not appreciably delay DNA synthesis in progress. Return to optimal temperature (28°C) 50 or 100 min later resulted in initiation of S, in delayed cells, at a rate greater than in controls. During the synchronization treatment, most cells were unable to enter S during a heat shock, but initiated S with a slight delay during the following intershock period. These cells were not appreciably delayed in completion of S by subsequent heat shocks. Supraoptimal temperature appears to affect the DNA synthetic cycle near the G1 to S transition. Cells subjected to the heat-shock treatment in early G1 all participated in one S period, and many underwent a succession of two S periods. DNA synthesis occurred in about 50% of the cells between EST and the first synchronous division, with the likelihood of DNA synthesis becoming greater the longer the interval between these two events. In some cells no detectable DNA synthesis occurred between EST and the second synchronous division. It was concluded that a precise temporal alternation of DNA replication and cell division is not obligatory in Tetrahymena.

scholarly journals P transposons controlled by the heat shock promoter.

1986 ◽  
Vol 6 (5) ◽  
pp. 1640-1649 ◽  
Author(s):  
H Steller ◽  
V Pirrotta
Keyword(s):  
Heat Shock ◽  
Germ Line ◽  
Fusion Gene ◽  
P Element ◽  
Translational Efficiency ◽  
Heat Shock Promoter ◽  
P Elements ◽  
Heat Shocks ◽  
Neomycin Resistance ◽  
Heat Shock Induction

We have transformed Drosophila melanogaster with modified P-element transposons, which express the transposase function from the heat-inducible hsp70 heat shock promoter. The Icarus transposon, which contains a direct hsp70-P fusion gene, behaved like a very active autonomous P element even before heat shock induction. Although heat shock led to abundant somatic transcription, transposition of the Icarus element was confined to germ line cells. To reduce the constitutive transposase activity observed for the Icarus element, we attenuated the translational efficiency of the transposase RNA by inserting the transposon 5 neomycin resistance gene between the hsp70 promoter and the P-element sequences. The resulting construct, called Icarus-neo, conferred resistance to G418, and its transposition was significantly stimulated by heat shock. Heat shocks applied during the embryonic or third instar larval stage had similar effects, indicating that transposition of P elements is not restricted to a certain developmental stage. Both Icarus and Icarus-neo destabilized snw in a P-cytotype background and thus at least partially overcome the repression of transposition. Our results suggest that the regulation of P-element transposition occurs at both the transcriptional and posttranscriptional levels.

scholarly journals Stable Formation of Compositionally Unique Stress Granules in Virus-Infected Cells

2009 ◽  
Vol 84 (7) ◽  
pp. 3654-3665 ◽  
Author(s):  
Joanna Piotrowska ◽  
Spencer J. Hansen ◽  
Nogi Park ◽  
Katarzyna Jamka ◽  
Peter Sarnow ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Heat Shock ◽  
Viral Infections ◽  
De Novo ◽  
Stress Granules ◽  
Stress Granule ◽  
Initiation Factor ◽  
Granule Formation ◽  
Infected Cells ◽  
Positive Strand Rna

ABSTRACT Stress granules are sites of mRNA storage formed in response to a variety of stresses, including viral infections. Here, the mechanisms and consequences of stress granule formation during poliovirus infection were examined. The results indicate that stress granules containing T-cell-restricted intracellular antigen 1 (TIA-1) and mRNA are stably constituted in infected cells despite lacking intact RasGAP SH3-domain binding protein 1 (G3BP) and eukaryotic initiation factor 4G. Fluorescent in situ hybridization revealed that stress granules in infected cells do not contain significant amounts of viral positive-strand RNA. Infection does not prevent stress granule formation in response to heat shock, indicating that poliovirus does not block de novo stress granule formation. A mutant TIA-1 protein that prevents stress granule formation during oxidative stress also prevents formation in infected cells. However, stress granule formation during infection is more dependent upon ongoing transcription than is formation during oxidative stress or heat shock. Furthermore, Sam68 is recruited to stress granules in infected cells but not to stress granules formed in response to oxidative stress or heat shock. These results demonstrate that stress granule formation in poliovirus-infected cells utilizes a transcription-dependent pathway that results in the appearance of stable, compositionally unique stress granules.

Seed germination response of a potential rangeland weed Psilocaulon granulicaule to selected environmental conditions

2020 ◽  
Vol 68 (5) ◽  
pp. 363
Author(s):  
Rekha Ranaweera ◽  
Sandra L. Weller ◽  
Singarayer K. Florentine
Keyword(s):  
Water Stress ◽  
Heat Shock ◽  
Seed Germination ◽  
Management Practices ◽  
Germination Rate ◽  
Radiant Heat ◽  
Weed Species ◽  
Light Regimes ◽  
Heat Shocks ◽  
Environmental Weed

Studies show that just over 620 non-native naturalised plant species have been recorded within the Australian rangelands, some of which have a capacity to cause significant impacts on rangeland flora and grazing activity. Although Psilocaulon granulicaule (Haw.), Schwantes is listed as a highly invasive environmental weed species, there has been no previous research into its seed ecology. Therefore, this study was conducted to investigate the effects of temperature, light, pH, water stress, heat-shock, and salinity on the germination of P. granulicaule. In this study, four temperature regimes covering four different day and night temperature variations (17–7°C, 25–15°C, 30–25°C and 40–30°C) and two light regimes (12-h light–12-h dark, 24-h dark) were investigated. The effects of pH, water stress, heat-shock and salinity were investigated, using pH buffers, polyethylene glycol solutions, three heat shock events under four temperatures and a range of NaCl solutions. These tests were conducted under the identified optimum temperature range (25/15°C) and light regime for seed germination. The results showed that both temperature and photoperiod significantly influenced the germination rate, with 94.2% germination in the 25–15°C range under a 12-h light-12-h dark regime. Higher temperatures (30–40°C) reduced seed germination to <58% germination in both light regimes (57.5%, 12-h light-12-h dark; 54.17%, 24-h dark). The highest germination rates were observed in low pH solutions, high moisture levels, low heat-shocks and low salinity. The study showed that this species is sensitive to environmental factors such as temperature, light, pH, moisture, heat shock and salinity, suggesting that these factors can be used as critical indicators to guide effective management practices to address this weed problem. Given that seeds are sensitive to radiant heat, burning could be used as a tool to effectively manage this species.

scholarly journals Repression of hsp70 heat shock gene transcription by the suppressor of hairy-wing protein of Drosophila melanogaster.

1991 ◽  
Vol 11 (4) ◽  
pp. 1894-1900 ◽  
Author(s):  
C Holdridge ◽  
D Dorsett
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Protein Binding ◽  
Protein Interactions ◽  
Binding Sites ◽  
Zinc Finger Protein ◽  
Gene Promoter ◽  
Heat Shock Gene ◽  
Protein Binding Sites ◽  
Shock Gene

The suppressor of hairy-wing [su(Hw)] locus of Drosophila melanogaster encodes a zinc finger protein that binds a repeated motif in the gypsy retroposon. Mutations of su(Hw) suppress the phenotypes associated with mutations caused by gypsy insertions. To examine the mechanisms by which su(Hw) alters gene expression, a fragment of gypsy containing multiple su(Hw) protein-binding sites was inserted into various locations in the well-characterized Drosophila hsp70 heat shock gene promoter. We found no evidence for activation of basal hsp70 transcription by su(Hw) protein in cultured Drosophila cells but observed that it can repress heat shock-induced transcription. Repression occurred only when su(Hw) protein-binding sites were positioned between binding sites for proteins required for heat shock transcription. We propose that su(Hw) protein interferes nonspecifically with protein-protein interactions required for heat shock transcription, perhaps sterically, or by altering the ability of DNA to bend or twist.

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