Heat Shock Conditions Research Articles

Characterizing Three Heat Shock Protein 70 Genes of Aphis gossypii and Their Expression in Response to Temperature and Insecticide Stress.

Aphis gossypii is a highly polyphagous pest that causes substantial agricultural damage. Temperature and insecticides are two major abiotic stresses affecting their population abundance. Heat shock proteins play an essential role in cell protection when insects are exposed to environmental stresses. Three ApHsp70 genes were cloned from A. gossypii, and characterized their molecular features and expression profiles in response to temperature and insecticide stress. The deduced amino acid sequences of these proteins exhibited characteristic Hsp70 family signatures, and their tissue-specific expression patterns revealed their highest activity to be in the salivary glands under 35 °C. The temperature inductive assay further indicated that the expression of the three ApHsp70 genes was markedly upregulated under heat stress but not under cold shock. Furthermore, exposure to LC25 and LC50 concentrations of three insecticides triggered the upregulation of these ApHsp70 genes. The RNA interference (RNAi)-mediated suppression of ApHsp68 expression heightened cotton aphid's susceptibility to insecticides (acetamiprid and sulfoxaflor). Moreover, our study found that the sulfoxaflor-resistant strain of A. gossypii (Sul-R) displayed a higher survival rate compared with the sulfoxaflor-sensitive strain (Sul-S) under heat shock conditions. These results suggest that these three ApHsp70 genes play an essential role in response to both heat and insecticide stress.

Hsf1 is essential for proteotoxic stress response in smyd1b-deficient embryos and fish survival under heat shock.

Molecular chaperones play critical roles in post-translational maintenance in protein homeostasis. Previous studies have shown that loss of Smyd1b function results in defective myofibril organization and dramatic upregulation of heat shock protein gene (hsp) expression in muscle cells of zebrafish embryos. To investigate the molecular mechanisms and functional importance of this stress response, we characterized changes of gene expression in smyd1b knockdown and knockout embryos using RNA-seq. The results showed that the top upregulated genes encode mostly cytosolic heat shock proteins. Co-IP assay revealed that the upregulated cytosolic Hsp70s associate with myosin chaperone UNC45b which is critical for myosin protein folding and sarcomere assembly. Strikingly, several hsp70 genes also display muscle-specific upregulation in response to heat shock-induced stress in zebrafish embryos. To investigate the regulation of hsp gene upregulation and its functional significance in muscle cells, we generated heat shock factor 1 (hsf-/-) knockout zebrafish mutants and analyzed hsp gene expression and muscle phenotype in the smyd1b-/-single and hsf1-/-;smyd1b-/- double-mutant embryos. The results showed that knockout of hsf1 blocked the hsp gene upregulation and worsened the muscle defects in smyd1b-/- mutant embryos. Moreover, we demonstrated that Hsf1 is essential for fish survival under heat shock (HS) conditions. Together, these studies uncover a correlation between Smyd1b deficiency and the Hsf1-activated heat shock response (HSR) in regulating muscle protein homeostasis and myofibril assembly and demonstrate that the Hsf1-mediated hsp gene upregulation is vital for the survival of zebrafish larvae under thermal stress conditions.

Elucidating stress responses in Prunus rootstocks through comprehensive evaluation under drought, heat shock and combined stress conditions

Elucidating stress responses in Prunus rootstocks through comprehensive evaluation under drought, heat shock and combined stress conditions

Bioinformatic Analysis of Actin-Binding Proteins in the Nucleolus During Heat Shock.

Actin plays a crucial role not only in the cytoplasm, but also in the nucleus, influencing various cellular behaviors, including cell migration and gene expression. Recent studies reveal that nuclear actin dynamics is altered by cellular stresses, such as DNA damage; however, the effect of heat shock on nuclear actin dynamics, particularly in the nucleolus, remains unclear. This study aims to elucidate the contribution of nucleolar actin to cellular responses under heat shock conditions. Nuclear actin dynamics in response to heat shock were investigated using nAC-GFP, a GFP-tagged actin chromobody, to visualize nuclear actin in HeLa cells. Bioinformatic analyses were also performed. Heat shock induced the reversible assembly of nAC-GFP in the nucleolus, with disassembly occurring upon recovery in a heat shock protein (Hsp) 70-dependent manner. Because the nucleolus, formed via liquid-liquid phase separation (LLPS), sequesters misfolded proteins under heat shock to prevent irreversible aggregation, we hypothesized that nucleolar actin-binding proteins might also be sequestered in a similar manner. Using several databases, we identified 47 actin-binding proteins localized in the nucleolus and determined the proportion of intrinsically disordered regions (IDRs) known to promote LLPS. Our analysis revealed that many of these 47 proteins exhibited high levels of IDRs. The findings from our bioinformatics analysis and further cellular studies may help elucidate new roles for actin in the heat shock response.

Adverse effects of heat shock in rice (Oryza sativa L.) and approaches to mitigate it for sustainable rice production under the changing climate: A comprehensive review.

Heat shock, a transient exposure to high temperatures, is a substantial hazard to rice (Oryza sativa L.) production and sustainability. The objective of this review paper is to summarize the impact of heat shock on rice and explore approaches to mitigate its adverse effects to achieve sustainable production. Rice is a staple food for billions of people globally and is extremely sensitive to heat shock. Higher temperatures disturb various physiological and biochemical processes, resulting in decreased growth, development, and ultimately lower grain yield. Heat shock negatively affects important agronomic traits, such as panicle differentiation, pollen viability, fertilization, grain filling, and, ultimately, grain quality. To manage heat shock and sustain rice production, several strategies have been explored, such as modifications to sowing schedules, the substitution of heat-tolerant cultivars for sensitive genotypes, and the use of growth regulators. To improve rice under heat shock, various approaches could be taken: (1) cultivating cultivars that flower early in the morning by adjusting sowing/planting times, modified irrigation, and fertilization; (2) inducing acclimation via growth regulators and organic stimulants and chemicals; (3) breeding genetically resistant cultivars through the integration of appropriate genes; and (4) genetic modification techniques for heat-shock tolerance. Overall, effectively managing heat-shock stress in rice requires a comprehensive strategy that includes developing and using heat shock-tolerant cultivars, adopting suitable cultural practices, utilizing external substances, and applying biotechnological tools. Implementing these strategies collectively will help achieve sustainable rice production in the face of increasing heat-shock conditions.

Unveiling the novel regulatory roles of RpoD-family sigma factors in Salmonella Typhimurium heat shock response through systems biology approaches.

Three RpoD-family sigma factors, RpoD, RpoS, and RpoH, play critical roles in transcriptional regulation in Salmonella enterica serovar Typhimurium under heat shock conditions. However, the genome-wide regulatory mechanisms of these sigma factors in response to heat stress have remained elusive. In this study, we comprehensively identified 2,319, 2,226, and 213 genome-wide binding sites for RpoD, RpoS, and RpoH, respectively, under sublethal heat shock conditions (42°C). Machine learning-based transcriptome analysis was employed to infer the relative activity of iModulons, providing valuable insights into the transcriptional impact of heat shock. Integrative data analysis enabled the reconstruction of the transcriptional regulatory network of sigma factors, revealing how they modulate gene expression to adapt to heat stress, including responses to anaerobic and oxidative stresses. Notably, we observed a significant expansion of the RpoS sigmulon from 97 to 301 genes in response to heat shock, underscoring the crucial role of RpoS in regulating various metabolic processes. Moreover, we uncovered a competition mechanism between RpoD and RpoS within RpoS sigmulons, where RpoS significantly increases its binding within promoter regions shared with RpoD under heat shock conditions. These findings illuminate how three RpoD-family sigma factors coordinate multiple cellular processes to orchestrate the overall response of S. Typhimurium to heat stress.

Mild Heat Stress Alters the Physical State and Structure of Membranes in Triacylglycerol-Deficient Fission Yeast, Schizosaccharomyces pombe.

We investigated whether the elimination of two major enzymes responsible for triacylglycerol synthesis altered the structure and physical state of organelle membranes under mild heat shock conditions in the fission yeast, Schizosaccharomyces pombe. Our study revealed that key intracellular membrane structures, lipid droplets, vacuoles, the mitochondrial network, and the cortical endoplasmic reticulum were all affected in mutant fission yeast cells under mild heat shock but not under normal growth conditions. We also obtained direct evidence that triacylglycerol-deficient cells were less capable than wild-type cells of adjusting their membrane physical properties during thermal stress. The production of thermoprotective molecules, such as HSP16 and trehalose, was reduced in the mutant strain. These findings suggest that an intact system of triacylglycerol metabolism significantly contributes to membrane protection during heat stress.

Entamoeba histolytica: In Silico and In Vitro Oligomerization of EhHSTF5 Enhances Its Binding to the HSE of the EhPgp5 Gene Promoter.

Throughout its lifecycle, Entamoeba histolytica encounters a variety of stressful conditions. This parasite possesses Heat Shock Response Elements (HSEs) which are crucial for regulating the expression of various genes, aiding in its adaptation and survival. These HSEs are regulated by Heat Shock Transcription Factors (EhHSTFs). Our research has identified seven such factors in the parasite, designated as EhHSTF1 through to EhHSTF7. Significantly, under heat shock conditions and in the presence of the antiamoebic compound emetine, EhHSTF5, EhHSTF6, and EhHSTF7 show overexpression, highlighting their essential role in gene response to these stressors. Currently, only EhHSTF7 has been confirmed to recognize the HSE as a promoter of the EhPgp5 gene (HSE_EhPgp5), leaving the binding potential of the other EhHSTFs to HSEs yet to be explored. Consequently, our study aimed to examine, both in vitro and in silico, the oligomerization, and binding capabilities of the recombinant EhHSTF5 protein (rEhHSTF5) to HSE_EhPgp5. The in vitro results indicate that the oligomerization of rEhHSTF5 is concentration-dependent, with its dimeric conformation showing a higher affinity for HSE_EhPgp5 than its monomeric state. In silico analysis suggests that the alpha 3 α-helix (α3-helix) of the DNA-binding domain (DBD5) of EhHSTF5 is crucial in binding to the major groove of HSE, primarily through hydrogen bonding and salt-bridge interactions. In summary, our results highlight the importance of oligomerization in enhancing the affinity of rEhHSTF5 for HSE_EhPgp5 and demonstrate its ability to specifically recognize structural motifs within HSE_EhPgp5. These insights significantly contribute to our understanding of one of the potential molecular mechanisms employed by this parasite to efficiently respond to various stressors, thereby enabling successful adaptation and survival within its host environment.

Glutathione S-Transferase Genes Involved in Response to Short-Term Heat Stress in Tetranychus urticae (Koch).

Tetranychus urticae, a globally ubiquitous mite, poses a significant threat to agriculture. Elevated temperatures exacerbate the growth, development, and reproduction of T. urticae, leading to substantial crop damage. In this study, we employed comparative transcriptomic approaches with whole-genome information of T. urticae to identify six Glutathione S-transferase genes (GSTs) implicated in heat stress response. Through comprehensive bioinformatics analyses, we elucidated the tertiary structure and active sites of the corresponding proteins, providing a thorough characterization of these GST genes. Furthermore, we investigated the expression patterns of these six GST genes under short-term heat shock conditions. Our findings unveiled the involvement of T. urticae GST genes in combating oxidative stress induced by heat, underscoring their role in antioxidant defense mechanisms. This study contributes valuable insights into the molecular mechanisms underlying the response of T. urticae to heat stress, laying a foundation for the development of strategies aimed at mitigating its impact in high-temperature environments.

The role of VdSti1 in Verticillium dahliae: insights into pathogenicity and stress responses.

Sti1/Hop, a stress-induced co-chaperone protein, serves as a crucial link between Hsp70 and Hsp90 during cellular stress responses. Despite its importance in stress defense mechanisms, the biological role of Sti1 in Verticillium dahliae, a destructive fungal pathogen, remains largely unexplored. This study focused on identifying and characterizing Sti1 homologues in V. dahliae by comparing them to those found in Saccharomyces cerevisiae. The results indicated that the VdSti1-deficient mutant displayed increased sensitivity to drugs targeting the ergosterol synthesis pathway, leading to a notable inhibition of ergosterol biosynthesis. Moreover, the mutant exhibited reduced production of microsclerotia and melanin, accompanied by decreased expression of microsclerotia and melanin-related genes VDH1, Vayg1, and VaflM. Additionally, the mutant's conidia showed more severe damage under heat shock conditions and displayed growth defects under various stressors such as temperature, SDS, and CR stress, as well as increased sensitivity to H2O2, while osmotic stress did not impact its growth. Importantly, the VdSti1-deficient mutant demonstrated significantly diminished pathogenicity compared to the wild-type strain. This study sheds light on the functional conservation and divergence of Sti1 homologues in fungal biology and underscores the critical role of VdSti1 in microsclerotia development, stress response, and pathogenicity of V. dahliae.

Contrasting responses of Thermocyclops crassus and T. oithonoides (Crustacea, Copepoda) to thermal stress.

Thermal tolerance is a critical factor influencing the survival of living organisms. This study focuses on the thermal resistance of copepod species, Thermocyclops crassus (Fischer, 1853) and T. oithonoides (Sars G.O., 1863), with overlapping distribution ranges in Europe. Short-term heat shock experiments were conducted to assess the thermal resistance of these copepods, considering various temperature increments and exposure durations. Additionally, the study explored the influence of heat shock on egg sac shedding, a vital indicator of population dynamics. Results indicate that widely distributed T. crassus exhibits higher thermal tolerance compared to narrowly distributed T. oithonoides, with survival rates varying under different heat shock conditions. Furthermore, T. crassus demonstrated a quicker response in dropping egg sacs in response to thermal stress, suggesting a potential adaptive mechanism for the survival of adults. However, rapid egg sac droppings pose high risks for eggs facing unfavorable conditions. T. crassus, inhabiting environments with greater temperature fluctuations such as the littoral and pelagial zones, exhibited better survival mechanisms compared to T. oithonoides, which predominantly resides in the pelagic zone. The findings have implications for understanding copepod responses to global warming and thermal pollution. This research contributes insights into the adaptive strategies of thermophilic copepod species and their ecological consequences.

Oxidative stress changes the effectiveness of artemisinin in Plasmodium falciparum.

Emerging resistance to the frontline antimalarial drug artemisinin represents a significant threat to worldwide malaria control and elimination. The patterns of parasite changes associated with emerging resistance represent a complex array of metabolic processes evident in various genetic mutations and altered transcription profiles. Genetic factors identified in regulating P. falciparum sensitivity to artemisinin overlap with the parasite's responses to malarial fever, sickle trait, and other types of oxidative stresses, suggesting conserved inducible survival responses. In this study we show that intraerythrocytic stress conditions, oxidative stress and heat shock, can significantly decrease the sensitivity of the parasite to artemisinin and lumefantrine, respectively. These results indicate that an intraerythrocytic oxidative stress microenvironment and heat-shock condition can alter antimalarial drug efficacy. Evaluating efficacy of antimalarial drugs under ideal in vitro culture conditions may not accurately predict drug efficacy in all malaria patients.

Calycosin Enhances Heat Shock Related-Proteins in H9c2 Cells to Modulate Survival and Apoptosis against Heat Shock.

Heat shock proteins (HSPs), which function as chaperones, are activated in response to various environmental stressors. In addition to their role in diverse aspects of protein production, HSPs protect against harmful protein-related stressors. Calycosin exhibits numerous beneficial properties. This study aims to explore the protective effects of calycosin in the heart under heat shock and determine its underlying mechanism. H9c2 cells, western blot, TUNEL staining, flow cytometry, and immunofluorescence staining were used. The time-dependent effects of heat shock analyzed using western blot revealed increased HSP expression for up to 2[Formula: see text]h, followed by protein degradation after 4[Formula: see text]h. Hence, a heat shock damage duration of 4[Formula: see text]h was chosen for subsequent investigations. Calycosin administered post-heat shock demonstrated dose-dependent recovery of cell viability. Under heat shock conditions, calycosin prevented the apoptosis of H9c2 cells by upregulating HSPs, suppressing p-JNK, enhancing Bcl-2 activation, and inhibiting cleaved caspase 3. Calycosin also inhibited Fas/FasL expression and activated cell survival markers (p-PI3K, p-ERK, p-Akt), indicating their cytoprotective properties through PI3K/Akt activation and JNK inhibition. TUNEL staining and flow cytometry confirmed that calycosin reduced apoptosis. Moreover, calycosin reversed the inhibitory effects of quercetin on HSF1 and Hsp70 expression, illustrating its role in enhancing Hsp70 expression through HSF1 activation during heat shock. Immunofluorescence staining demonstrated HSF1 translocation to the nucleus following calycosin treatment, emphasizing its cytoprotective effects. In conclusion, calycosin exhibits pronounced protective effects against heat shock-induced damages by modulating HSP expression and regulating key signaling pathways to promote cell survival in H9c2 cells.

Dendritic Cells Loaded With Heat Shock Inactivated Glioma Stem Cells Enhance Antitumor Response of Mouse Glioma When Combining With CD47 Blockade.

For glioma patients, the long-term advantages of dendritic cells (DCs) immunization remain unknown. It is extremely important to develop new treatment strategies that enhance the immunotherapy effect of DC-based vaccines. DCs exposed to glioma stem cells (GSCs) are considered promising vaccines against glioma. Glioma stem cells were isolated from mouse glioma GL261 cells (GCs). Both were subjected to severe (47°C) and mild (42°C) heat shock to induce immunogenic cell death (ICD). Membrane mobilization of calreticulin (CRT) and release of heat shock proteins (HSPs) were detected by flow cytometry. Dendritic cells were then exposed to heat-inactivated cells and co-culturing of T cells tested for immunotherapeutic efficacy in vitro. In vivo, we investigated the GSC targeting effect of the GSC-DC vaccine combined with CD47 blockade. Heat shock induced ICD in GCs and GSCs, as indicated by significant release of calreticulin, HSP70, and HSP90. Heat shock condition ICD lysates induce maturation and activation-associated marker expression on monocyte-derived DCs. Accordingly, DCs pulsed with GCs and GSCs inactivated reduced colony formation, sphere formation, migration, and invasion of glioma and GSCs in vitro. Glioma stem cell-DC vaccine in combination with anti-CD47 antibody significantly enhanced survival in mice with glioma, induced production of interferon (IFN)-γ, and enhanced T-cell expansion in vivo. Of note, DCs pulsed with inactivated GSCs were more effective to control tumor growth than DCs pulsed with inactive GCs. Severe heat shock induces ICD in vitro. These data showed that administration of anti-CD47 antibody combined with GSC-DC vaccine may represent an effective immunotherapeutic strategy for cancer patients in clinical.

Assessment of physiological and biochemical thermotolerance traits in tomato genotypes

The increasing frequency and duration of high-temperature extremes have adversely impacted tomato production globally. An improved understanding of the physio-biochemical traits under such climatic changes will enable the selection, development, and adoption of resilient tomato varieties. Thus, this study investigated a suite of fifteen tomato plant traits that could be involved in the physio-biochemical mechanisms of thermotolerance to facilitate the selection of heat-tolerant tomato varieties. Ten tomato varieties selected from a previously published study were exposed to three temperature conditions (26/18 °C as control, 38/28 °C for 7 d as heat stress, and 40 °C for 7 h as heat-shock treatment). The results demonstrated that a composite variable of membrane injury metrics, including electrolyte leakage, malondialdehyde content, and heat injury index, was the most successful variable for discriminating varietal thermotolerance under either heat stress or heat shock conditions among all the evaluated traits. Principal components of other plant physiological traits performed poorly in distinguishing thermotolerance among the varieties and essentially did not match the results from the membrane injury composite variable. We identified ‘Celebrity,’ ‘Heat Master,’ and ‘Valley Girl’ as the most thermotolerant varieties under both heat treatment conditions, whereas ‘Arkansas Traveler’, ‘Pruden's Purple’, and ‘Tasti-Lee’ were found to be the most thermosensitive. These results echoed results from a previous open-field screening trial of 43 tomato varieties. Results from this study suggest that measurements of membrane injury are more effective than leaf physiological traits and that heat shock treatments may be as effective as longer term, moderate heat stress treatments for screening tomato thermotolerance, implying that future controlled environment screenings may reduce the number of variables measured and the length of their trials.

The ability to induce heat shock transcription factor-regulated genes in response to lethal heat stress is associated with thermotolerance in tomato cultivars.

Heat stress is a severe challenge for plant production, and the use of thermotolerant cultivars is critical to ensure stable production in high-temperature-prone environments. However, the selection of thermotolerant cultivars is difficult due to the complex nature of heat stress and the time and space needed for evaluation. In this study, we characterized genome-wide differences in gene expression between thermotolerant and thermosensitive tomato cultivars and examined the possibility of selecting gene expression markers to estimate thermotolerance among different tomato cultivars. We selected one thermotolerant and one thermosensitive cultivar based on physiological evaluations and compared heat-responsive gene expression in these cultivars under stepwise heat stress and acute heat shock conditions. Transcriptomic analyses reveled that two heat-inducible gene expression pathways, controlled by the heat shock element (HSE) and the evening element (EE), respectively, presented different responses depending on heat stress conditions. HSE-regulated gene expression was induced under both conditions, while EE-regulated gene expression was only induced under gradual heat stress conditions in both cultivars. Furthermore, HSE-regulated genes showed higher expression in the thermotolerant cultivar than the sensitive cultivar under acute heat shock conditions. Then, candidate expression biomarker genes were selected based on the transcriptome data, and the usefulness of these candidate genes was validated in five cultivars. This study shows that the thermotolerance of tomato is correlated with its ability to maintain the heat shock response (HSR) under acute severe heat shock conditions. Furthermore, it raises the possibility that the robustness of the HSR under severe heat stress can be used as an indicator to evaluate the thermotolerance of crop cultivars.

Plasmodium falciparum GCN5 plays a key role in regulating artemisinin resistance-related stress responses.

Plasmodium falciparum causes the most severe malaria and is exposed to various environmental and physiological stresses in the human host. Given that GCN5 plays a critical role in regulating stress responses in model organisms, we aimed to elucidate PfGCN5's function in stress responses in P. falciparum. The protein level of PfGCN5 was substantially induced under three stress conditions [heat shock, low glucose starvation, and dihydroartemisinin, the active metabolite of artemisinin (ART)]. With a TetR-DOZI conditional knockdown (KD) system, we successfully down-regulated PfGCN5 to ~50% and found that KD parasites became more sensitive to all three stress conditions. Transcriptomic analysis via RNA-seq identified ~1,000 up- and down-regulated genes in the wild-type (WT) and KD parasites under these stress conditions. Importantly, DHA induced transcriptional alteration of many genes involved in many aspects of stress responses, which were heavily shared among the altered genes under heat shock and low glucose conditions, including ART-resistance-related genes such as K13 and coronin. Based on the expression pattern between WT and KD parasites under three stress conditions, ~300-400 genes were identified to be involved in PfGCN5-dependent, general, and stress-condition-specific responses with high levels of overlaps among three stress conditions. Notably, using ring-stage survival assay, we found that KD or inhibition of PfGCN5 could sensitize the ART-resistant parasites to the DHA treatment. All these indicate that PfGCN5 is pivotal in regulating general and ART-resistance-related stress responses in malaria parasites, implicating PfGCN5 as a potential target for malaria intervention.

Molecular, enzymatic responses and in vitro embryonic developmental competency of heat-shocked buffalo embryos co-cultured with granulosa cells monolayer

The present study was designed to establish a suitable alternative approach to mitigate the adverse effect of high culture temperature on in vitro embryo development and the related molecular response in buffalo. Pre-cultured granulosa cells (GCs) were used as a monolayer during in vitro embryo culture until day 8 (day of fertilization = D0). Post fertilization, presumptive embryos were randomly assigned into two culture conditions: embryos cultured in the presence of GCs monolayer under normal culture temperature (N: 38.5 °C; GEN group) or heat shock (H: 40.5 °C; GEH group) and their counterpart groups of embryos cultured without GCs (EN and EH groups). Additionally, two groups of GCs monolayer were cultured without embryos up to day 8 under 38.5 °C (GN) or 40.5 °C (GH) for further spent culture media enzymatic analyses. Heat shock was administered for the first 2 h of culture then continued at 38.5 °C until day 8. The results indicated that under heat treatment, GCs enhanced (P ≤ 0.05) embryo cleavage and development (day 8) rates, which were comparable to the embryos cultured at 38.5 °C. On the molecular level, blastocysts of the GEH group showed similar expressions of metabolism-regulating genes (CPT2 and SlC2A1/GLUT1) and an antioxidant gene (SOD2) when compared to the blastocysts of the EN group. The relative expression of HSP90 was significantly up-regulated under heat shock and/or co-culture conditions. However, HSF1 expression was increased (P ≤ 0.05) in the GEH group. No statistical differences were observed among the study groups for the pluripotency gene NANOG, and stress resistance transcript NFE2L2. Regarding the enzymatic profile, the concentrations of SOD, total protein, and MDA were decreased (P ≤ 0.05) in the GEH group compared to the cultured GCs without embryos (GH group). In conclusion, GCs as a monolayer have a beneficial impact on alleviating heat stress at the zygote stage through the regulatory mechanisms of metabolic activity, defense system, and heat shock response genes.

Functional characterization of the DNA-binding protein from starved cells (DPS) as a molecular chaperone under heat stress

The DNA-binding protein from starved cells, known as DPS, has been characterized as a crucial factor in protecting E. coli from external stresses. The DPS functions in various cellular processes, including protein-DNA binding, ferroxidase activity, compaction of chromosome and regulation of stress resistance gene expression. DPS proteins exist as oligomeric complexes; however, the specific biochemical activity of oligomeric DPS in conferring heat shock tolerance has not been fully understood. Therefore, we investigated the novel functional role of DPS under heat shock. To elucidate the functional role of DPS under heat shock conditions, we purified recombinant GST-DPS protein and demonstrated its thermostability and existence in its highly oligomeric form. Furthermore, we discovered that the hydrophobic region of GST-DPS influenced the formation of oligomers, which exhibited molecular chaperone activity, thereby preventing the aggregation of substrate proteins. Collectively, our findings highlight the novel functional role of DPS, as a molecular chaperone and may confer thermotolerance to E. coli.

Screening of Different Media and Heat Shock Treatment Regimens for Enhancing Sporulation in Bacillus licheniformis

Bacillus licheniformis is a spore-producing bacteria. The present study aims to identify maximum sporulation in Bacillus licheniformis through induced heat shock conditions using different solidified media and broth media. Suspensions of Bacillus licheniformis spores were spread-plated on nutrient agar plates before and after heat shock treatments of 60°C, 80°C and 100°C for 30 minutes. The number of spores was determined by the dilution plate count method. The result indicates that the highest spore production for Bacillus licheniformis was observed when the spores were induced in AK (Arret and Kirshbaum) Agar for seven days under a heat shock treatment of 60℃ for 30 minutes. Whereas, in the case of Nutrient Broth media, the spore production was highest after four days at heat shock treatment of 80℃ for 30 minutes. This study will help find the optimal production of spores from Bacillus licheniformis, an industrially beneficial microorganism.