scholarly journals Expression of heat shock protein and trehalose-6-phosphate synthase homologues induced during water deficit in cotton

2002 ◽  
Vol 14 (1) ◽  
pp. 11-20 ◽  
Author(s):  
Alexandre Lima Nepomuceno ◽  
Derrick Oosterhuis ◽  
James McD Stewart ◽  
Rickie Turley ◽  
Norman Neumaier ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Water Deficit ◽  
Protein Denaturation ◽  
Differentially Expressed ◽  
Water Deficit Stress ◽  
Cdna Clones ◽  
Drought Period ◽  
Cellular Dehydration ◽  
Phosphate Synthase

Tolerance to drought in plants is not a simple trait, but a complex of mechanisms working in combination to avoid or to resist water deficit. Genotypes that differ in tolerance to water deficit may show qualitative and quantitative differences in gene expression when submitted to drought periods. Four cotton (Gossypium hirsutum L.) genotypes (Siokra L-23, Stoneville 506, CS 50 and T-1521) with contrasting responses to water deficit stress were studied using the Differential Display (DD) technique to identify and isolate genes which may differ among them. Fifty-two cDNA fragments differentially expressed during water deficit were isolated, cloned and sequenced. Search of gene bank databases showed that two cDNA clones, A12B15-6 and A12B13-1, have high homology with a heat shock protein that binds to calmodulin found in Nicotiana tabacum (2.9e-32 P(N)) and with an Arabidopsis thaliana trehalose-6-phosphate synthase enzyme (9.0e-37 P(N)), respectively. One of the presumed functions of heat shock proteins is related to prevention of protein denaturation during cellular dehydration. Trehalose-6-phosphate synthase is involved in the production of trehalose, a disaccharide known to osmotically protect cell membranes during dehydration. The HSP homologue was found to be differentially expressed during the drought period in two drought tolerant genotypes but not in drought-sensitive genotypes. The trehalose-6-phosphate synthase homologue was also up-regulated during water deficit stress, however, all four genotypes were induced to express this homologue. Ribonuclease protection assays confirmed these results. This is an important finding since there are only few reports of trehalose presence in higher plants and none in cotton.

scholarly journals Genomewide analysis of circular RNA in pituitaries of normal and heat-stressed sows

BMC Genomics ◽  
2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Haojie Zhang ◽  
Baoyu Hu ◽  
Jiali Xiong ◽  
Ting Chen ◽  
Qianyun Xi ◽  
...  
Keyword(s):  
Heat Stress ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Noncoding Rna ◽  
Target Genes ◽  
Hormone Secretion ◽  
Circular Rna ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Pituitary Hormone

Abstract Background As a newly characterized type of noncoding RNA, circular RNA (circRNA) has been shown to have functions in diverse biological processes of animals. It has been reported that several noncoding RNAs may regulate animals’ response to heat stress which can be easily induced by hyperthermia in summer. However, the expression and functions of circRNAs in the pituitary of sows and whether they participate in heat stress adaption are still unclear. Results In this study, we found that high temperature over the thermoneutral zone of sows during the summer increased the serum heat shock protein 70 (HSP70) level, decreased the superoxide dismutase (SOD) vitality and prolactin (PRL) concentration, and induced heat stress in sows. Then, we explored circRNA in the pituitary of heat-stressed and normal sows using RNA sequencing and bioinformatics analysis. In total, 12,035 circRNAs were detected, with 59 circRNAs differentially expressed, including 42 up-regulated and 17 down-regulated circRNAs in pituitaries of the heat-stressed sows. Six randomly selected circRNAs were identified through reverse transcription PCR followed by DNA sequencing and other 7 randomly selected differentially expressed circRNAs were verified by quantitative real-time PCR analysis. The predicted target genes regulated by circRNAs through sponging microRNAs (miRNAs) were enriched in metabolic pathway. Furthermore, the predicted circRNA–miRNA–mRNA interactions showed that some circRNAs might sponge miRNAs to regulate pituitary-specific genes and heat shock protein family members, indicating circRNA’s roles in pituitary hormone secretion and heat stress response. Conclusions Our results provided a meaningful reference to understand the functions of circRNA in the porcine pituitary and the mechanisms by which circRNA may participate in animals’ response to heat stress.

scholarly journals Transcriptome Functional Analysis of Mammary Gland of Cows in Heat Stress and Thermoneutral Condition

Animals ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1015 ◽  
Author(s):  
Shuangming Yue ◽  
Zhisheng Wang ◽  
Lizhi Wang ◽  
Quanhui Peng ◽  
Bai Xue
Keyword(s):  
Functional Analysis ◽  
Immune Response ◽  
Heat Stress ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Milk Production ◽  
Milk Yield ◽  
Molecular Mechanisms ◽  
Mammary Glands ◽  
Differentially Expressed

Heat stress (HS) exerts significant effects on the production of dairy animals through impairing health and biological functions. However, the molecular mechanisms related to the effect of HS on dairy cow milk production are still largely unknown. The present study employed an RNA-sequencing approach to explore the molecular mechanisms associated with a decline in milk production by the functional analysis of differentially expressed genes (DEGs) in mammary glands of cows exposed to HS and non-heat-stressed cows. The results of the current study reveal that HS increases the rectal temperature and respiratory rate. Cows under HS result in decreased bodyweight, dry matter intake (DMI), and milk yield. In the current study, a total of 213 genes in experimental cow mammary glands was identified as being differentially expressed by DEGs analysis. Among identified genes, 89 were upregulated, and 124 were downregulated. Gene Ontology functional analysis found that biological processes, such as immune response, chaperone-dependent refolding of protein, and heat shock protein binding activity, were notably affected by HS. The Kyoto Encyclopedia of Genes and Genomes enrichment analysis found that almost all of the top-affected pathways were related to immune response. Under HS, the expression of heat shock protein 90 kDa beta I (HSP90B1) and heat shock 70 kDa protein 1A was upregulated, while the expression of bovine lymphocyte antigen (BoLA) and histocompatibility complex, class II, DRB3 (BoLA-DRB3) was downregulated. We further explored the effects of HS on lactation-related genes and pathways and found that HS significantly downregulated the casein genes. Furthermore, HS increased the expression of phosphorylation of mammalian target of rapamycin, cytosolic arginine sensor for mTORC1 subunit 2 (CASTOR2), and cytosolic arginine sensor for mTORC1 subunit 1 (CASTOR1), but decreased the phosphorylation of Janus kinase-2, a signal transducer and activator of transcription factor-5. Based on the findings of DMI, milk yield, casein gene expression, and the genes and pathways identified by functional annotation analysis, it is concluded that HS adversely affects the immune function of dairy cows. These results will be beneficial to understand the underlying mechanism of reduced milk yield in HS cows.

scholarly journals Identification of proteins differentially expressed by glutamate treatment in cerebral cortex of neonatal rats

2019 ◽  
Vol 35 (1) ◽  
Author(s):  
Ju-Bin Kang ◽  
Dong-Ju Park ◽  
Phil-Ok Koh
Keyword(s):  
Cerebral Cortex ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Cell Damage ◽  
Neuronal Cell ◽  
Differentially Expressed ◽  
Heat Shock Protein 60 ◽  
Sprague Dawley ◽  
Two Dimensional Gel Electrophoresis ◽  
Specific Proteins

AbstractGlutamate leads to neuronal cell damage by generating neurotoxicity during brain development. The objective of this study is to identify proteins that differently expressed by glutamate treatment in neonatal cerebral cortex. Sprague-Dawley rat pups (post-natal day 7) were intraperitoneally injected with vehicle or glutamate (10 mg/kg). Brain tissues were isolated 4 h after drug treatment and fixed for morphological study. Moreover, cerebral cortices were collected for protein study. Two-dimensional gel electrophoresis and mass spectrometry were carried out to identify specific proteins. We observed severe histopathological changes in glutamate-exposed cerebral cortex. We identified various proteins that differentially expressed by glutamate exposure. Identified proteins were thioredoxin, peroxiredoxin 5, ubiquitin carboxy-terminal hydrolase L1, proteasome subunit alpha proteins, isocitrate dehydrogenase, and heat shock protein 60. Heat shock protein 60 was increased in glutamate exposed condition. However, other proteins were decreased in glutamate-treated animals. These proteins are related to anti-oxidant, protein degradation, metabolism, signal transduction, and anti-apoptotic function. Thus, our findings can suggest that glutamate leads to neonatal cerebral cortex damage by regulation of specific proteins that mediated with various functions.

Proteomic Analysis Identifies Differences in Multiple Myeloma (MM) Cells Sensitive and Resistant to AKT Inhibition.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3402-3402
Author(s):  
Alissa Huston ◽  
Alexey Leontovich ◽  
Michael Timm ◽  
Yazan Alsayed ◽  
Ujjal Singha ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Cell Lines ◽  
Proteomic Analysis ◽  
Molecular Mechanisms ◽  
Treatment Time ◽  
Control Sample ◽  
Differentially Expressed ◽  
Akt Inhibitor ◽  
The Mean

Abstract Prior studies have demonstrated that MM cells with PTEN mutation and high AKT activity are more sensitive to inhibitors of the PI3K/Akt/mTOR pathway. However, the molecular mechanisms that regulate the differential response to these agents are not well characterized. The objective of this study was to determine proteins that are different between MM cell lines with higher AKT activity (OPM2) and lower AKT activity (MM.1S) in response to the AKT inhibitor, perifosine. Methods: MM cell lines (MM.1S and OPM2) were treated with serial concentration of perifosine (KRX-0401, Keryx, NY, NY, provided by the NCI). Proteomic analysis using the nanoscale BD Clontech antibody-based protein microarray technique was performed using cells treated with perifosine (10uM for 16 hrs) or vehicle (sterile water) as control. Apoptosis was determined using Annexin V/PI FACS analysis at 24 and 48 hrs. The treatment time and concentration were chosen so that it did not induce more than 25% apoptosis to ensure adequate analysis of changes in signaling pathways. The antibody microarray is a technique that detects differences in protein abundance between the treated and control sample with each experiment by hybridizing fluorescently labeled (Cy3 and Cy5) protein mixtures onto slides spotted with 512 human monoclonal antibodies. Two microarray slides were used for each experiment. The slides were scanned using the Axon GenePix 4000B scanner. Two ratios were generated from the spot images for each protein target. The mean of the ratios of Cy5/Cy3 of both slides were analyzed using Clontech software and used to calculate an Internally Normalized Ratio (INR = (Ratio1/Ratio2, ratios 1 and 2 correspond to slides 1 and 2) for each spot on the array. The INR values were input into GeneSpring 6.0 software (Silicon Genetics, CA). The data was normalized to the mean INR of the two cell lines. Proteins whose expression level changed relative to control greater than 1.3 fold were determined. Unsupervised clustering demonstrated a different protein signature between MM.1S and OPM2 in response to perifosine. There were 144 proteins differentially expressed by 1.3 fold between MM.1S and OPM2. Proteins that were downregulated in OPM2 as compared to MM.1S included those in the PI3K pathway and cell cycle regulation such as PTEN, p70S6Kinase, the AKT substrate GSK-3, eEF-2 kinase, eIF-4g, Ku-80, cyclin A, E2F-2, CDK2, CDK7, and c-myc; proteins involved in apoptosis such as p21WAF, caspase 4 and 8, FADD, and PARP; kinases such as PKAc, PKA RI, ERK2, and JNK1; and other proteins regulating apoptosis and proliferation including p53, the NF-kB inhibitor IkB, and the heat shock protein HSP70. The nanoscale protein array is a useful and rapid technique that may be used to identify differences between resistant and sensitive cells to novel therapeutic agents. We identified proteins that are differentially expressed between MM cells sensitive and relatively resistant to the AKT inhibitor perifosine. Further analysis of the role of these proteins in the mechanism of resistance/sensitivity to perifosine is being performed. Future use of inhibitors of NF-kB (bortezomib) or heat shock protein inhibitors in conjunction with perifosine may overcome resistance induced by these proteins in MM cells with low AKT activity. Supported in part by an ASH scholar award and an MMRF grant.

Proteomics Response and Acclimation of Camptotheca acuminata Seedlings to Water Deficit

2012 ◽  
Vol 518-523 ◽  
pp. 460-464
Author(s):  
Jing Hua Yu ◽  
Shu Sheng Yuan ◽  
Zhong Hua Tang ◽  
De Wen Li ◽  
Yuan Gang Zu
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Water Deficit ◽  
Arid Environment ◽  
Camptotheca Acuminata ◽  
Abundant Protein ◽  
Drought Conditions ◽  
Camptotheca Acuminata Seedlings ◽  
Identification Analysis ◽  
Late Embryo

Proteomics responses and adaptations of Camptotheca acuminata seedlings to drought conditions stimulated by treatment with PEG8000 simulation were investigated. We determined the drought responses of seedlings after 30min, 3 h, and 5 h of treatment with15% PEG8000 . The following 2-DE and PMF identification analysis showed that there are many kinds of proteins involved in the regulation of plants responses to environmental drought. Heat shock protein (HSP) and Late embryo abundant protein (LEA) were discovered to take part in the response of C. acuminata to drought environment. Rubisco LSU was found to help C. acuminata to adapt this arid environment in the way of degradation.

scholarly journals Heat shock protein 70 and glycoprotein 96 are differentially expressed on the surface of malignant and nonmalignant breast cells

2006 ◽  
Vol 11 (4) ◽  
pp. 334 ◽  
Author(s):  
Karla Melendez ◽  
Erik S. Wallen ◽  
Bruce S. Edwards ◽  
Charlotte D. Mobarak ◽  
David G. Bear ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Heat Shock Protein 70 ◽  
Differentially Expressed ◽  
Breast Cells

scholarly journals Human homologues of the bacterial heat-shock protein DnaJ are preferentially expressed in neurons

1992 ◽  
Vol 284 (2) ◽  
pp. 469-476 ◽  
Author(s):  
M E Cheetham ◽  
J P Brion ◽  
B H Anderton
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Human Brain ◽  
Atp Hydrolysis ◽  
Sequence Similarity ◽  
Polyclonal Antiserum ◽  
Cdna Clones ◽  
Expression Library ◽  
Dnaj Protein ◽  
Protein Complex Dissociation

The bacterial heat-shock protein DnaJ has been implicated in protein folding and protein complex dissociation. The DnaJ protein interacts with the prokaryotic analogue of Hsp70, DnaK, and accelerates the rate of ATP hydrolysis by DnaK. Several yeast homologues of DnaJ, with different proposed subcellular localizations and functions, have recently been isolated and are the only eukaryotic forms of DnaJ so far described. We have isolated cDNAs corresponding to two alternatively spliced transcripts of a novel human gene, HSJ1, which show sequence similarity to the bacterial DnaJ protein and the yeast homologues. The cDNA clones were isolated from a human brain-frontal-cortex expression library screened with a polyclonal antiserum raised to paired-helical-filament (PHF) proteins isolated from extracts of the brains of patients suffering from Alzheimer's disease. The similarity between the predicted human protein sequences and the bacterial and yeast proteins is highest at the N-termini, this region also shows a limited similarity to viral T-antigens and is a possible common motif involved in the interaction with DnaK/Hsp70. Northern-blot analysis has shown that human brain contains higher levels of mRNA for the DnaJ homologue than other tissues examined, and hybridization studies with riboprobes in situ show a restricted pattern of expression of the mRNA within the brain, with neuronal layers giving the strongest signal. These findings suggest that the DnaJ-DnaK (Hsp70) interaction is general to eukaryotes and, indeed, to higher organisms.

34 DIFFERENTIALLY EXPRESSED PROTEINS OF EARLY-STAGE PLACENTA DERIVED FROM CLONED CAT EMBRYOS USING PROTEOMICS ANALYSIS

2012 ◽  
Vol 24 (1) ◽  
pp. 129
Author(s):  
J. I. Bang ◽  
D. W. Bae ◽  
Y. S. Kwon ◽  
G. K. Deb ◽  
B. H. Choi ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Early Pregnancy ◽  
Protein Disulfide Isomerase ◽  
Triosephosphate Isomerase ◽  
Early Stage ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Disulfide Isomerase ◽  
Protein Disulfide

We have previously demonstrated that differentially expressed proteins affect abnormal development and function of cloned term placenta. This is associated with cloned fetus morbidity and mortality. We also frequently observed loss of the cloned fetus and failed development during early pregnancy periods. To confirm the pattern of important gene expression in cloned placenta during pre- and post-implantation, we investigated expression pattern of proteins in early stage (21 days) domestic cat placentas of cloned embryo transfer (CEP; n = 2) and artificial insemination (CP; n = 4) derived pregnancy. The differentially expressed proteins were investigated by 2-DE and MALDI-TOF/MS. Twenty-three proteins were up- and down-regulated at least 1.5-fold in the CEP (P < 0.05) compared with the CP. Differentially expressed proteins were analysed using PDQest program and statistically analysed by 1-way ANOVA using the SPSS software. In CEP, 13 proteins were up-regulated, such as 78-kDa glucose-regulated protein (GRP78), annexin A2 (ANXA2), protein DJ-1 (DJ1), adenylate kinase isoenzyme 1 (AK1), protein disulfide-isomerase A3 (PDIA3), heat shock protein β-1 (HSPB1), actin, cytoplasmic 1 (ACTB), serum albumin (ALB), protein disulfide-isomerase A6 (PDIA6), G protein-regulated inducer of neurite outgrowth 1 (GRIN1) and triosephosphate isomerase (TIM). In contrast, 10 proteins were down-regulated, such as vinculin (VCL), triosephosphate isomerase (TIM), heterogeneous nuclear ribonucleoprotein H (hnRNPH), tropomyosin α-4 (TPM4), 60-kDa heat shock protein, mitochondrial (Hsp60), serum albumin (ALB), calumenin (CALU), keratin type 1 (CK1) and prohibitin (PHB). To validate the identified proteins in the CEP compared with the CP, we investigate a peptide sequences using MALDI-TOF/TOF tandem mass spectrometry. The sequence information obtained a high ions score from NCBI and Swiss-Prot databases. In conclusion, we did identify abnormal expression of proteins that might be associated with impaired development of CEP, which may endanger the cloned fetus during early pregnancy. This work was partly supported by the BK21 program and the KOSEF (10525010001-05N2501-00110) and the Next-generation BioGreen21 program (No. PJ007990012011).

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