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1.
The mitochondrial 60-kDa heat shock protein in marine invertebrates: biochemical purification and molecular characterization
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Sessile marine invertebrates undergo constant direct exposure to the surrounding environmental conditions, including local and global environmental fluctuations that may lead to fatal protein damage. Induction of heat shock proteins (Hsps) constitutes an important defense mechanism that protects these organisms from deleterious stress conditions. In a previous study, we reported the immunological detection of a 60-kDa Hsp (Hsp60) in the sea anemone Anemonia viridis (formerly called Anemonia sulcata) and studied its expression under a variety of stress conditions. In the present study, we show that the sponge Tetilla sp. from tidal habitats with a highly variable temperature regime is characterized by an increased level of Hsp60. Moreover, we show the expression of Hsp60 in various species among Porifera and Cnidaria, suggesting a general importance of this protein among marine invertebrates. We further cloned the hsp60 gene from A viridis, using a combination of conventional protein isolation methods and screening of a complementary deoxyribonucleic acid library by polymerase chain reaction. The cloned sequence (1764 bp) encodes for a protein of 62.8 kDa (588 amino acids). The 62.8-kDa protein, which contains an amino terminal extension that may serve as a mitochondrial targeting signal, shares a significant identity with mitochondrial Hsp60s from several animals but less identity with Hsp60s from either bacteria or plants. 相似文献
2.
Jianke Yang Juan Gu Yuqing Hu Nan Wang Jiguang Gao Ping Wang 《Cell stress & chaperones》2021,26(1):115
Heat shock protein 60 (HSP60) is a well-recognized multifunctional protein, playing a substantial role in protecting organisms from environmental stress. The domestic pigeon (Columba livia) is a promising model organism, with important economic and ecological value, and its health is susceptible to temperature stress. To explore the molecular characteristics, tissue expression profile, and response to temperature stress for HSP60 of Columba livia (ClHSP60), we firstly cloned and characterized the complete cDNA sequence and investigated its expression profile under optimal conditions and acute temperature stress. The cDNA of ClHSP60 contained 2257 nucleotides, consisting of 12 exons with length ranging from 65 to 590 bp. The open reading frame (ORF) encoded 573 amino acids with calculated molecular weight of 60.97 kDa that contained a number of structurally prominent domains or motifs. Under optimal temperature conditions, levels of ClHSP60 expression differed between all the tested tissues (the highest was noted in liver and the lowest in pectoralis major muscle). Under acute temperature stress, five patterns of change were detected in the tested tissues, suggesting that different tissues in domestic pigeons differentially responded to various temperature stress conditions. Upregulation of ClHSP60 expression was highest in the lung and pectoralis major muscle, reflecting the crucial role of these two tissues in temperature regulation. However, the crop, cerebrum, and heart showed little change or decreased ClHSP60 expression. The results indicate that ClHSP60 may be sensitive to and play pivotal roles in responding to acute temperature stress.Electronic supplementary materialThe online version of this article (10.1007/s12192-020-01160-7) contains supplementary material, which is available to authorized users. 相似文献
3.
Jianfeng Ding Jia Li Dongmin Yang Feng Yang Hongtao Nie Zhongming Huo Xiwu Yan 《Cell stress & chaperones》2018,23(2):179-187
4.
Organisms produce stress proteins as a response to natural and anthropogenic environmental changes. Induction of stress proteins has been reported in a variety of aquatic organisms, including rotifers, exposed to pollutants. Past studies on stress protein responses of rotifers have focused on exposure to single toxicants. In this study the rotifer Plationus patulus was exposed singly and in combination to various concentrations of As, Cr, Cu, Ni, Pb, and Zn. Following exposure, total protein was quantified (Bradford method) and stress protein 60 (HSP60) was identified using Western blotting. P. patulus induced HSP60 as a response to single exposures to Cr, Cu, Ni, Pb and Zn. HSP60 expression was increased (2 fold) in rotifers exposed to these single elements at both low and high concentrations as compared to unexposed rotifers. Arsenic exposure resulted in a 2 fold decrease in HSP induction. In rotifers exposed to metal mixtures, HSP60 was induced by the presence of As–Zn, As–Cr–Cu–Pb, As–Cr–Cu, As–Cr–Cu–Ni and As–Cr–Cu–Ni–Pb combinations in the media. HSP60 response to As and heavy metals toxicity depends on the type and number of elements present in the media as well as their concentrations and length of the exposure time. 相似文献
5.
In the yeast Saccharomyces cerevisiae, the molecular chaperone HSP26 has the remarkable ability to sense increases in temperature directly and can switch from
an inactive to a chaperone-active state. In this report, we analyzed the effect of expression of HSP26 in Arabidopsis thaliana plants and their response to high temperature stress. The hsp26 transgenic plants exhibited stronger growth than wild type plants at 45 °C for 16 h. The chlorophyll content and chlorophyll
fluorescence decreased much more in wild type than in transgenic plants. Moreover, the transgenic plants had higher proline
and soluble sugar contents, and lower relative electrical conductivity and malondialdehyde contents after high temperature
stress. Furthermore, we found that over-expression of HSP26 in Arabidopsis increased the amount of free proline, elevated the expression of proline biosynthetic pathway genes and therefore enhanced
Arabidopsis tolerance to heat stress. 相似文献
6.
7.
Zahida Zahoor Angela J. Davies Ruth S. Kirk David Rollinson Anthony John Walker 《Cell stress & chaperones》2010,15(5):639-650
Synthesis of heat shock proteins (HSPs) following cellular stress is a response shared by many organisms. Amongst the HSP
family, the ∼70 kDa HSPs are the most evolutionarily conserved with intracellular chaperone and extracellular immunoregulatory
functions. This study focused on the effects of larval excretory-secretory products (ESPs) from the parasite Schistosoma mansoni on HSP70 protein expression levels in haemocytes (defence cells) from its snail intermediate host Biomphalaria glabrata. S. mansoni larval stage ESPs are known to interfere with haemocyte physiology and behaviour. Haemocytes from two different B. glabrata strains, one which is susceptible to S. mansoni infection and one which is resistant, both showed reduced HSP70 protein levels following 1 h challenge with S. mansoni ESPs when compared to unchallenged controls; however, the reduction observed in the resistant strain was less marked. The
decline in intracellular HSP70 protein persisted for at least 5 h in resistant snail haemocytes only. Furthermore, in schistosome-susceptible snails infected by S. mansoni for 35 days, haemocytes possessed approximately 70% less HSP70. The proteasome inhibitor, MG132, partially restored HSP70
protein levels in ESP-challenged haemocytes, demonstrating that the decrease in HSP70 was in part due to intracellular degradation.
The extracellular signal-regulated kinase (ERK) signalling pathway appears to regulate HSP70 protein expression in these cells,
as the mitogen-activated protein-ERK kinase 1/2 (MEK1/2) inhibitor, U0126, significantly reduced HSP70 protein levels. Disruption
of intracellular HSP70 protein expression in B. glabrata haemocytes by S. mansoni ESPs may be a strategy employed by the parasite to manipulate the immune response of the intermediate snail host. 相似文献
8.
In the present paper we suggest an effect of seasonal variations in food availability on two ecophysiological parameters in
a warm temperate benthic suspension feeder: the tissue concentrations of proteins, carbohydrates and lipids on the one hand,
and the expression of stress proteins (HSP 70 and 90, inducible and/or constitutive) on the other hand. The concentrations
of biomacromolecules have already been used to describe bentho-pelagic and reproductive processes, but this is the first time
that stress protein expression is suggested to be directly related with food constraints in marine organisms. Paramuricea clavata (Cnidaria: Gorgonacea) express HSP 70 and 90 (constitutive and/or inducible) throughout the seasonal cycle, and HSP 70 levels
are twice as high as the levels of HSP 90. In summer and autumn, when seston availability to suspension feeders was low, P. clavata showed low levels of carbohydrates and lipids, but high levels of HSPs expression. The levels of HSP 70 and 90 expression
fit with negative exponential functions of carbohydrate and lipid concentrations. We suggest a direct effect of food availability
on the studied ecophysiological parameters while the effect of temperature may be rather indirect. HSP expression as well
as the tissue concentrations of carbohydrate and lipids may be used as biomarkers of environmental changes and seston availability
to benthic suspension feeders. 相似文献
9.
Heat shock proteins (HSPs) are a family of genes classically used to measure levels of organism stress. We have previously
identified two HSP70 genes (HSP70A and HSP70B) in sub-tidal populations of the Antarctic limpet (Nacella concinna). These genes are up-regulated in response to increased seawater temperatures of 15°C or more during acute heat shock experiments,
temperatures that have very little basis when considering the current Antarctic ecology of these animals. Therefore, the question
was posed as to whether these animals could express HSP70 genes when subjected to more complex environmental conditions, such
as those that occur in the inter-tidal. Inter-tidal limpets were collected on three occasions in different weather conditions
at South Cove, Rothera Point, over a complete tidal cycle, and the expression levels of the HSP70 genes were measured. Both
genes showed relative up-regulation of gene expression over the period of the tidal cycle. The average foot temperature of
these animals was 3.3°C, far below that of the acute heat shock experiments. These experiments demonstrate that the temperature
and expression levels of HSP production in wild animals cannot be accurately extrapolated from experimentally induced treatments,
especially when considering the complexity of stressors in the natural environment. However, experimental manipulation can
provide molecular markers for identifying stress in Antarctic molluscs, provided it is accompanied by environmental validation,
as demonstrated here.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
10.
K Osman I Ibrahim A Yousef T Nabil A Nayerah 《World journal of microbiology & biotechnology》2012,28(5):1997-2001
Bacterial heat-shock response is a global regulatory system required for effective adaptation to changes (stress) in the environment.
An in vitro study was conducted to investigate the impact of a sublethal temperature (42°C) on heat shock protein (HSP) expression
in 6 Salmonella strains (Salmonella Enteritidis, S. Typhimurium, S. Virchow, S. Shubra, S. Haifa and S. Eingedi). The 6 Salmonella strains were isolated from the tissues of ducklings that had died from avian salmonellosis. To
determine the induction of HSP in the 6 Salmonella strains, they were exposed to the selected temperature level for 24 h and further kept for 48 h at culturing condition of
42°C. Growth under a sublethal temperature of 42°C increased the expression of several proteins of Salmonella, including a 63 kDa protein in addition to the generation and/or overexpression of 143 proteins which were specific to heat
shock, concurrent to this acquired thermotolerance. The 6 Salmonella strains responded to 24 h of thermal stress at an elevated temperature 42°C by synthesizing different heat shock proteins
(HSP) with molecular weights ranging between 13.62 and 96.61 kDa. At 48 h, the 6 Salmonella strains synthesized different HSPs with molecular weights ranging between 14.53 and 103.43 kDa. It follows that salmonellae
would produce HSPs during the course of the infectious process. Salmonellosis produced several proteins after 24 and 48 h
of infection. Seven of these proteins (100, 80, 60, 40, 30, 20 and 10 kDa) were recognized in the serum obtained from the
ducklings infected with S. Enteritidis, S. Typhimurium, S. Virchow, S. Shubra, S. Haifa and S. Eingedi after 24 h of infection. After 48 h, the 1–7 kDa HSP became more evident and indicated their de novo generation. 相似文献
11.
Uttarani Maibam O. K. Hooda Ph. Suraj Sharma S. V. Singh A. K. Mohanty R. C. Upadhyay 《Biological Rhythm Research》2017,48(4):647-661
The study aimed to evaluate inducible HSP70 (HSP70.1 and HSP70.2) gene expression and oxidative stress status in skin of cattle during different seasons. Ten each of Tharparkar (zebu) and Karan Fries (crossbred) heifers were selected from NDRI herd, Karnal. Animals were maintained under standard managemental practices followed at the farm. Skin biopsies were aseptically collected from each animal during winter, spring, and summer. Real time PCR was performed to examine HSP70 expression. Reactive oxygen species (ROS) and antioxidant enzymes (SOD and CAT) were determined by ELISA. In both the breeds, significantly higher (p < 0.05) levels of HSP70 expression, ROS, caspases, and antioxidant enzymes were observed during summer followed by winter and spring. Breeds showed no significant difference during winter and spring. During summer, HSP70 expression, ROS, and antioxidant enzymes were higher (p < 0.05) in Karan Fries than Tharparkar, whereas caspases levels were higher in Tharparker than Karan Fries. The study concludes that levels of HSP70 expression, ROS, caspases, and antioxidant enzymes in skin of cattle were strongly affected by seasonal change in temperature. Differences exist in skin tissue thermotolerance of Tharparkar and Karan Fries cattle. This might be an efficient and centrally important mechanism for better adaptability of zebu cattle to heat stress. 相似文献
12.
The Antarctic limpet, Nacella concinna, exhibits the classical heat shock response, with up-regulation of duplicated forms of the inducible heat shock protein 70
(HSP70) gene in response to experimental manipulation of seawater temperatures. However, this response only occurs in the
laboratory at temperatures well in excess of any experienced in the field. Subsequent environmental sampling of inter-tidal
animals also showed up-regulation of these genes, but at temperature thresholds much lower than those required to elicit a
response in the laboratory. It was hypothesised that this was a reflection of the complexity of the stresses encountered in
the inter-tidal region. Here, we describe a further series of experiments comprising both laboratory manipulation and environmental
sampling of N. concinna. We investigate the expression of HSP70 gene family members (HSP70A, HSP70B, GRP78 and HSC70) in response to a further suite
of environmental stressors: seasonal and experimental cold, freshwater, desiccation, chronic heat and periodic emersion. Lowered
temperatures (−1.9°C and −1.6°C), generally produced a down-regulation of all HSP70 family members, with some up-regulation
of HSC70 when emerging from the winter period and increasing sea temperatures. There was no significant response to freshwater
immersion. In response to acute and chronic heat treatments plus simulated tidal cycles, the data showed a clear pattern.
HSP70A showed a strong but very short-term response to heat whilst the duplicated HSP70B also showed heat to be a trigger,
but had a more sustained response to complex stresses. GRP78 expression indicates that it was acting as a generalised stress
response under the experimental conditions described here. HSC70 was the major chaperone invoked in response to long-term
stresses of varying types. These results provide intriguing clues not only to the complexity of HSP70 gene expression in response
to environmental change but also insights into the stress response of a non-model species. 相似文献
13.
Yong-Yu Li Shuai Lu Kun Li Jia-Yan Feng Yan-Na Li Zhi-Rong Gao Chang-Jie Chen 《Cell stress & chaperones》2010,15(6):965-975
The objective of this study was to investigate the function of heat shock protein 60 (HSP60) on pancreatic tissues by applying
HSP60 small interfering RNA (siRNA) to reduce HSP60 expression. Rat pancreas was isolated and pancreatic tissue snips were
prepared, cultured, and stimulated with low and high concentrations of cerulein (10−11 and 10−5 mol/L) or lipopolysaccharide (LPS, 10 and 20 μg/mL). Before the stimulation and 1 and 4 h after the stimulation, the viability
and the level of trypsinogen activation peptide (TAP) in the tissue fragments were determined and the levels of tumor necrosis
factor-alpha (TNF-α) and interleukin 6 (IL-6) in the culture supernatants were measured. Real-time PCR and Western blotting
were used to evaluate the HSP60 mRNA and protein expression. After the administration of siRNA to inhibit HSP60 expression
in the isolated tissues, these injury parameters were measured and compared. The pancreatic tissues in the control (mock-interfering)
group showed a decreased viability to varying degrees after being stimulated with cerulein or LPS, and the levels of TAP,
TNF-α, and IL-6 increased significantly (p < 0.05) in the tissues and/or in the culture supernatant. The expressions of HSP60 mRNA and protein were raised moderately after
stimulating 1 h with low concentrations of cerulein or LPS, but decreased with high concentrations of the toxicants. In particular,
the expression of HSP60 protein was reduced significantly (p < 0.05) when the tissues were stimulated by the two toxicants for 4 h. In contrast, the tissue fragments in which HSP60 siRNA
was applied showed much lower tissue viability (p < 0.01) and higher levels of TNF-a, IL-6, and TAP (p < 0.01) in the tissues or culture supernatant after stimulating with the toxicants at the same dose and for the same time duration
as compared with those of the control groups (p < 0.05). The results indicated that both cerulein and LPS can induce injuries on isolated pancreatic tissues, but the induction
effects are dependent on the duration of the stimulation and on the concentrations of the toxicants. HSP60 siRNA reduces HSP60
expression and worsens the cerulein- or LPS-induced injuries on isolated pancreatic tissues, suggesting that HSP60 has a protective
effect on pancreatic tissues against these toxicants. 相似文献
14.
Cara JB Aluru N Moyano FJ Vijayan MM 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》2005,142(4):426-431
Heat shock proteins (HSPs) expression is commonly used as indicators of cellular stress in animals. However, very little is known about either the expression patterns of HSPs or their role in the stress-tolerance phenomenon in early life stages of fish. To this end, we examined the impact of food-deprivation (12 h), reduced oxygen levels (3.5 mg/L for 1 h) and heat shock (HS: + 5 °C for 1 h) on HSP70 and HSP90 protein expression in early life stages of the gilthead sea bream (Sparus aurata), a warm-water aquaculture species. Also, we investigated HSP70 and HSP90 response to food-deprivation (7 days) in early life stages of rainbow trout (Oncorhynchus mykiss), a cool-water aquaculture species, and the tolerance of this larvae to heat shock (either + 5 or + 10 °C for 1 h). Our results clearly demonstrate that food-deprivation enhances HSP70 and HSP90 protein expression in larvae of both species. In gilthead sea bream larvae, the stressors-induced HSP70 and HSP90 (only in the reduced oxygen group) protein expression returned to unstressed levels after 24 h recovery. In fed trout larvae, a + 5 °C heat shock did not elevate HSP70 and HSP90 expression, whereas 100% mortality was evident with a + 10 °C HS. However, food-deprived trout larvae, which had higher HSP70 and HSP90 protein content, survived HS and showed HS-dependent increases in HSP70, but not HSP90 expression. Overall, HSP70 and HSP90 protein expression in early life stages of fish have the potential to be used as markers of nutritional stress, while elevation of the tissue HSPs content may be used as a means to increase stress tolerance during larval rearing. 相似文献
15.
16.
Yong-Yu Li Xue-Jin Li Shuai Lv Kun Li Yan-Na Li Zhi-Rong Gao Jia-Yan Feng Chang-Jie Chen Claus Schaefer 《Cell stress & chaperones》2010,15(5):583-591
Acute pancreatitis (AP) is an inflammatory process in which cytokines and chemokines are involved. After onset, extrapancreatic
stimuli can induce the expression of cytokines in pancreatic acinar cells, thereby amplifying this inflammatory loop. To further
determine the role and mechanism of irritating agents in the pathogenesis of AP, rat pancreatic tissues were stimulated with
ascitic fluid (APa) and serum (APs) from rats with AP or with lipopolysaccharide (LPS). In addition, the alteration of heat
shock protein 60 (HSP60) expression was evaluated. Rat pancreas was removed and meticulously snipped to fragments. The snips
were cultured for up to 48 h. During this period, the tissue viability as well as amylase and TNF-α levels in the supernatant
and the HSP60 expression in the pancreatic tissue before and after stimulation by APa, APs, and LPS were assayed time-dependently.
At different time-points during the culture, the viability and the amylase activity in the pancreatic tissue remained largely
stable. After stimulation with APa, APs, or LPS for 1 h, the pancreatic tissues showed some damage, and this was followed
by a sharp decrease in the viability accompanied by increased levels of amylase and TNF-α in the culture medium 2 or 4 h after
stimulation (p < 0.05). In contrast, both the HSP60 mRNA and protein levels had a relatively high expression in the freshly prepared tissue
fragments (0 h). As the culturing period was extended, the expression of HSP60 mRNA decreased only slightly; at the same time,
the HSP60 protein levels decreased over a prolonged culture time, significantly so from 12 through 48 h (p < 0.05). After stimulation with APs, APa, or LPS, both the expression of HSP60 mRNA and protein in the tissue fragments increased
slightly at 1 h and decreased significantly thereafter at 2 and 4 h (p < 0.05). APa, APs, or LPS induce injuries on isolated pancreatic tissues, accompanied by an altered HSP60 expression pattern
in a time-dependent manner. 相似文献
17.
Three AtHSP90 isoforms, cytosol-localized AtHSP90.2, chloroplast-localized AtHSP90.5, and endoplasmic reticulum (ER)-localized AtHSP90.7 genes, were constitutively overexpressed in Arabidopsis thaliana to study their functional mechanisms under oxidative stress. Overexpression of AtHSP90 genes reduced germination of transgenic seeds under oxidative stress. When exposed to 10 mM H2O2, AtHSP90 transgenic seedlings displayed lower activities of superoxide dismutase, catalase, and peroxidase; higher content of malondialdehyde;
and higher levels of protein damage than detected in the wild type. This indicated that overexpression of AtHSP90.2, AtHSP90.5, and AtHSP90.7 in Arabidopsis impaired plant tolerance to oxidative stress. Moreover, overexpression of chloroplast- and ER-localized AtHSP90 resulted in lower resistance to oxidative stress than that of cytosolic AtHSP90. This suggested that HSP90.2, HSP90.5, and HSP90.7 localized in different cellular compartments were involved in different functional mechanisms during oxidative stress. 相似文献
18.
Previous studies have shown that inhibiting the activity of the proteasome leads to the accumulation of damaged or unfolded
proteins within the cell. In this study, we report that proteasome inhibitors, lactacystin and carbobenzoxy-l-leucyl-l-leucyl-l-leucinal (MG132), induced the accumulation of ubiquitinated proteins as well as a dose- and time-dependent increase in the
relative levels of heat shock protein (HSP)30 and HSP70 and their respective mRNAs in Xenopus laevis A6 kidney epithelial cells. In A6 cells recovering from MG132 exposure, HSP30 and HSP70 levels were still elevated after
24 h but decreased substantially after 48 h. The activation of heat shock factor 1 (HSF1) may be involved in MG132-induced
hsp gene expression in A6 cells since KNK437, a HSF1 inhibitor, repressed the accumulation of HSP30 and HSP70. Exposing A6 cells
to simultaneous MG132 and mild heat shock enhanced the accumulation of HSP30 and HSP70 to a much greater extent than with
each stressor alone. Immunocytochemical studies determined that HSP30 was localized primarily in the cytoplasm of lactacystin-
or MG132-treated cells. In some cells treated with higher concentrations of MG132 or lactacystin, we observed in the cortical
cytoplasm (1) relatively large HSP30 staining structures, (2) colocalization of actin and HSP30, and (3) cytoplasmic areas
that were devoid of HSP30. Lastly, MG132 treatment of A6 cells conferred a state of thermotolerance such that they were able
to survive a subsequent thermal challenge. 相似文献
19.
Geoffrey E. Hill Xiaoyu Fu Susan Balenger Kevin J. McGraw Mathieu Giraudeau Wendy R. Hood 《Journal of Field Ornithology》2013,84(4):416-424
Heat‐shock proteins (HSP) are molecular chaperones that play key roles in the maintenance of cellular homeostasis under variable environmental conditions. Although HSP are frequently used in studies of wild vertebrates as indicators of stress, no one to date has assessed responses of HSP60, HSP70, and HSP90 in the same species to different environmental stressors. We studied changes in the circulating concentrations of HSP60, HSP70, and HSP90 in wild‐caught House Finches (Haemorhous mexicanus) in response to multiple and sequential stressful environments, including high temperatures, transportation, and pathogen exposure. House Finches sampled during a period of low‐environmental stress with moderate ambient temperatures had low levels of HSP60 and modest levels of HSP70 and HSP90 compared to birds sampled during a presumably more stressful period with high temperatures. After exposure to high‐ambient temperatures, transportation in a vehicle, and exposure to Mycoplasma gallisepticum, captive finches were found to have increasingly higher levels of HSP60. HSP70 tended to rise in response to each stressor, but to drop in the weeks between stress challenges. HSP90 levels increased significantly only in response to pathogen challenge. Our observations suggest that HSP60 and HSP70 are indices of a range of stressors in House Finches, whereas HSP90 primarily reflects health state. 相似文献