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1تقرير
المؤلفون: Ana Fernández, Ana Rita De Lima, Elizabeth Ferrer
مصطلحات موضوعية: Trypanosoma cruzi, heat shock protein, HSP83, cloning, expression
الوصف: [Objective] Tryapanosoma cruzi, causal agent of Chagas' disease, is a parasite that presents morphological alternation between an invertebrate (triatomine) and a vertebrate (mammal) host. Several studies have shown that in T. cruzi, HSP83 (homolog of HSP90) is essential for cell division and control of the response to thermal stress. The objetive of the present work was the cloning, bioinformatics characterization and expression of the heat shock protein HSP83 T. cruzi gene. [Methodology] RNA extraction was performed from epimastigotes using a commercial kit. The cDNA encoding HSP83 was obtained by RT-PCR, from the extracted mRNA, for which the primers were designed based on the HSP83 sequence of T. cruzi strain CL Brener. Cloning was performed on pGEM®T-Easy, and subcloned into the expression vector pQE30. Sequence and bioinformatics characterization was performed. The gene was expressed and the recombinant protein was purified by affinity chromatography and identify by immunoblotting. [Results] Sequence analysis showed similarity to the gene encoding HSP83 from Trypanosoma cruzi and HSP domains were observed, as well as B epitopes in the sequence. After 3 hours of induction with IPTG a recombinant protein with an approximate weight of 83 kDa was obtained. The immunoblotting reaction with hyperimmune anti-T. cruzi epimastigote serum allowed the detection a single band with a molecular weight of approximately 83 kDa [Conclusions]. All results indicate that the cloning and characterization of HSP83 from Trypanosoma cruzi was achieved.
العلاقة: https://zenodo.org/record/8252927Test; https://doi.org/10.5281/zenodo.8252927Test; oai:zenodo.org:8252927
الإتاحة: https://doi.org/10.5281/zenodo.8252927Test
https://doi.org/10.5281/zenodo.8252926Test
https://zenodo.org/record/8252927Test -
2تقرير
المؤلفون: Ciesielski, Szymon J, Schilke, Brenda A, Stolarska, Milena, Tonelli, Marco, Tomiczek, Bartlomiej, Craig, Elizabeth A
المصدر: FEBS Lett ; ISSN:1873-3468 ; Volume:598 ; Issue:12
مصطلحات موضوعية: Hsp40, JDP, J‐domain, heat shock protein, molecular chaperone, protein homeostasis
الوصف: J-domain proteins are critical Hsp70 co-chaperones. A and B types have a poorly understood glycine-rich region (Grich) adjacent to their N-terminal J-domain (Jdom). We analyzed the ability of Jdom/Grich segments of yeast Class B Sis1 and a suppressor variant of Class A, Ydj1, to rescue the inviability of sis1-∆. In each, we identified a cluster of Grich residues required for rescue. Both contain conserved hydrophobic and acidic residues and are predicted to form helices. While, as expected, the Sis1 segment docks on its J-domain, that of Ydj1 does not. However, data suggest both interact with Hsp70. We speculate that the Grich-Hsp70 interaction of Classes A and B J-domain proteins can fine tune the activity of Hsp70, thus being particularly important for the function of Class B.
العلاقة: https://doi.org/10.1002/1873-3468.14857Test; https://pubmed.ncbi.nlm.nih.gov/38529663Test; https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11209805Test/
الإتاحة: https://doi.org/10.1002/1873-3468.14857Test
https://pubmed.ncbi.nlm.nih.gov/38529663Test
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11209805Test/ -
3تقرير
مصطلحات موضوعية: ectotherms, general stress response, heat shock protein, stress tolerance, thermal stress
الوصف: Protein abundances used for all analyses.
الإتاحة: https://doi.org/10.5061/dryad.9km73/1Test
https://doi.org/10.5061/dryad.9km73Test
http://hdl.handle.net/10255/dryad.141363Test -
4تقرير
المؤلفون: Sun, Zhenjie, Gu, Wenhui, Feng, Zezhong, Fan, Yaqin, Niu, Jianfeng, Wang, Guangce
مصطلحات موضوعية: heat shock protein 70, high-temperature stress, AMP-activated protein kinase, cAMP-dependent protein kinase, Neopyropia yezoensis, Environmental Sciences & Ecology, Marine & Freshwater Biology, Environmental Sciences
الوصف: The upregulation of heat shock protein 70 (hsp70) gene under high temperature stress is a common phenomenon. Although heat shock protein-mediated stress responses play an important role in intertidal Neopyropia yezoensis, the detailed regulatory mechanism of the hsp70 gene expression is still unclear. Here, a full-length sequence of the hsp70-2 gene was cloned and its' expression regulation was analyzed. There was an activating transcription factors element (ATFE) of cAMP-dependent protein kinase (PKA) was found at the gene promoter region and a highly conserved deduced amino acid sequence with calmodulin-binding activity was detected. Reagents implicated in the induction of the hsp70 gene were then selected to treat the algal samples at 24 degrees C, and the photosynthetic parameters, transcription and translation of this gene were determined. Results showed that quercetin inhibited the transcription of the hsp70-2 gene, significantly decreased the synthesis of the HSP70-2 protein, and lowered the photosynthetic activity of N. yezoensis under high temperature stress conditions. Although the addition of trifluoperazine (TFP), an inhibitor of calmodulin (CAM), downregulated the photosynthetic parameters, the transcription of the hsp70-2 gene was not influenced at high temperature treatment, implying that CAM was not involved in the transcription of the hsp70-2 gene but involved in the heat stress reponding pathways. 5 '-aminoimidasole-4-carboxamide-1-beta-d-ribofuranoside (AICAR) promoted the synthesis of hsp70-2 transcripts significantly and HSP70-2 protein slightly, which indicated that AMPK regulated the induction of the hsp70-2 gene in N. yezoensis. Forskolin also promoted the upregulation of the hsp70-2 gene. Thus, both AMPK and cAMP-dependent protein kinase (PKA) could phosphorylate HSF and activate the expression of the hsp70-2 gene in N. yezoensis. However, there was no strict correlation between transcripts of the hsp70-2 gene and HSP70-2 protein contents. It was proposed that the posttranscriptional ...
العلاقة: FRONTIERS IN MARINE SCIENCE; http://ir.qdio.ac.cn/handle/337002/180554Test; http://ir.qdio.ac.cn/handle/337002/180555Test
الإتاحة: https://doi.org/10.3389/fmars.2022.1004600Test
http://ir.qdio.ac.cn/handle/337002/180554Test
http://ir.qdio.ac.cn/handle/337002/180555Test -
5تقرير
المؤلفون: Bayih, Abebe, Folefoc, Asongna, Mohon, Abu, Eagon, Scott, Anderson, Marc, Pillai, Dylan
مصطلحات موضوعية: Malaria, Plasmodium falciparum, Anti-malarial drugs, Heat-shock protein 90
الوصف: Background The emergence of artemisinin-resistant Plasmodium falciparum strains poses a serious challenge to the control of malaria. This necessitates the development of new anti-malarial drugs. Previous studies have shown that the natural beta-carboline alkaloid harmine is a promising anti-malarial agent targeting the P. falciparum heat-shock protein 90 (PfHsp90). The aim of this study was to test the anti-malarial activity of harmine analogues. Methods Forty-two harmine analogues were synthesized and the binding of these analogues to P. falciparum heat shock protein 90 was investigated. The in vitro anti-malarial activity of two of the analogues, 17A and 21A, was evaluated using a 72-h growth inhibition assay. The in vivo anti-malarial activity was tested in Plasmodium berghei infection of BALB/c mice. The potential of 21A for a combination treatment with artemisinin was evaluated using in vivo combination study with dihydro-artemisinin in BALB/c mice. Cytotoxicity of the harmine analogues was tested in vitro using HepG2 and HeLa cell lines. Results 17A and 21A bound to PfHsp90 with average IC 50 values of 12.2 ± 2.3 and 23.1 ± 8.8 µM, respectively. They also inhibited the P. falciparum W2 strain with average IC 50 values of 4.2 ± 1.3 and 5.7 ± 1.7 µM, respectively. In vivo, three daily injections of P. berghei -infected BALB/c mice with 100 mg/kg of either 17A or 21A showed significant reduction in parasitaemia with a 51.5 and 56.1% reduction, respectively. Mice treated with 17A and 21A showed a median survival time of 11 and 14 days, respectively, while the vehicle control mice survived a median of only 8.5 days. A dose-ranging experiment with 21A showed that the compound has a dose-dependent anti-malarial effect. Furthermore, treatment of infected mice with a combination of 21A and dihydroartemisinin (DHA) showed a dramatic reduction in parasitaemia compared to treatment with DHA alone. Conclusion A novel and non-toxic harmine analogue has been synthesized which binds to PfHsp90 protein, inhibits ...
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6تقرير
المؤلفون: Zou, Yan-Feng, Xu, Jian-Hua, Gu, Yuan-Yuan, Pan, Fa-Ming, Tao, Jin-Hui, Wang, De-Guang, Xu, Sheng-Qian, Xiao, Hui, Chen, Pei-Ling, Liu, Shuang, Cai, Jing, Lian, Li, Liu, Sheng-Xiu, Liang, Chun-Mei, Tian, Guo, Ye, Qian-Ling, Pan, Hai-Feng, Su, Hong, Ye, Dong-Qing
مصطلحات موضوعية: Systemic lupus erythematosus, Glucocorticoids, Heat shock protein 90, Single nucleotide polymorphisms
الوصف: Heat shock protein 90 (HSP90) is an important glucocorticoid receptor (GR) chaperone protein, and is supposed to be the key factor in regulating glucocorticoids (GCs) effects. The aim of the present study was to explore whether single nucleotide polymorphisms (SNPs) within HSP90AA1 gene affect the response of systemic lupus erythematosus (SLE) patients to GCs treatment. Two hundred and forty-five SLE patients were treated with GCs (prednisone) for 12 weeks. SLE disease activity index (SLEDAI) was used to assess the response of SLE patients to GCs treatment, and patients were classified into sensitive group and insensitive group. HapMap database and Haploview software were used to select tag SNPs. Tag SNPs were genotyped by using multiplex SNaPshot method. Univariate and multivariate logistic regression analyses were used to discriminate the impact of SNPs of HSP90AA1 gene on the response of SLE patients to GCs treatment. Two hundred and thirty three SLE patients finished the 12-week follow-up. Of these patients, 128 patients were included in sensitive group, and 105 patients were included in insensitive group. Seven tag SNPs were selected within HSP90AA1 gene. We detected significant associations for rs7160651 (dominant model: crude OR 0.514, 95 % CI 0.297–0.890, P = 0.018; adjusted OR 0.518, 95 % CI 0.293–0.916, P = 0.024), rs10873531 (dominant model: crude OR 0.516, 95 % CI 0.305–0.876, P = 0.014; adjusted OR 0.522, 95 % CI 0.304–0.898, P = 0.019) and rs2298877 (dominant model: crude OR 0.543, 95 % CI 0.317–0.928, P = 0.026, adjusted OR 0.558, 95 % CI 0.323–0.967, P = 0.037) polymorphisms, but not for other polymorphisms ( P > 0.05). The present study demonstrates that HSP90AA1 gene SNPs may affect the response of SLE patients to GCs treatment.
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7تقرير
المؤلفون: Li, Shuangjiang, Zhang, Wenbiao, Fan, Jun, Lai, Yutian, Che, Guowei
مصطلحات موضوعية: Heat shock protein 27, Non-small cell lung cancer, Prognosis, Meta-analysis
الوصف: Numbers of clinical and experimental investigations have provided increasing evidences to demonstrate that heat shock protein 27 (HSP27) is a qualified predictor for many cancers. However, no consensus has been reached on its clinicopathological and prognostic significance in patients with non-small cell lung cancer (NSCLC). Therefore, we performed this systematic meta-analysis to help addressing this issue. PubMed, EMBASE, the Web of Science and China National Knowledge Infrastructure were searched for full-text literatures met out eligibility criteria. We determined the odds ratio (OR) and hazard ratio (HR) as the appropriate summarized statistics for assessments of clinicopathological and prognostic roles of HSP27, respectively. Q-test and I 2 -statistic were used to evaluate the level of heterogeneity. Sensitivity analysis was conducted to examine the stability of overall estimates. Potential publication bias was detected by Begg’s test and Egger’s test. Finally, ten articles were identified to be included into our meta-analysis. The pooled analyses suggested that HSP27 expression was significantly associated with the unfavorable conditions for differentiation degree, lymphatic metastasis, clinical stage, squamous cell carcinoma and tumor size. However, HSP27 expression had no significant relationship to gender, age and smoking status. Meanwhile, pooled HRs indicated that HSP27 expression could be a predictor for a lower 5-year overall survival (OS) rate (HR: 1.832; 95 % CI 1.322–2.538; P < 0.001) but not for 1-year OS of NSCLC (HR: 0.885; 95 % CI 0.140–5.599; P = 0.896). In conclusion, our meta-analysis demonstrates that HSP27 expression may be a strong biomarker to predict both the poor clinicopathological and prognostic characteristics in patients with NSCLC.
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8تقريرModeling Alexander disease with patient iPSCs reveals cellular and molecular pathology of astrocytes
المؤلفون: Kondo, Takayuki, Funayama, Misato, Miyake, Michiyo, Tsukita, Kayoko, Era, Takumi, Osaka, Hitoshi, Ayaki, Takashi, Takahashi, Ryosuke, Inoue, Haruhisa
مصطلحات موضوعية: Alexander disease (AxD), Glial fibrillary acidic protein (GFAP), Induced pluripotent stem cells (iPSCs), Disease modeling, Astrocytes, Rosenthal fibers, Heat-shock protein, Alpha-crystallin, Cytokine, Inflammatory response, Inherited astrocytopathy
الوصف: Alexander disease is a fatal neurological illness characterized by white-matter degeneration and formation of Rosenthal fibers, which contain glial fibrillary acidic protein as astrocytic inclusion. Alexander disease is mainly caused by a gene mutation encoding glial fibrillary acidic protein, although the underlying pathomechanism remains unclear. We established induced pluripotent stem cells from Alexander disease patients, and differentiated induced pluripotent stem cells into astrocytes. Alexander disease patient astrocytes exhibited Rosenthal fiber-like structures, a key Alexander disease pathology, and increased inflammatory cytokine release compared to healthy control. These results suggested that Alexander disease astrocytes contribute to leukodystrophy and a variety of symptoms as an inflammatory source in the Alexander disease patient brain. Astrocytes, differentiated from induced pluripotent stem cells of Alexander disease, could be a cellular model for future translational medicine.
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9تقرير
المؤلفون: Li, Junyang, Tang, Chao, Li, Liwen, Li, Rujun, Fan, Youwu
مصطلحات موضوعية: Glioma, Soluble epoxide hydrolase, Heat shock protein 27, Cyclooxygenase 2, Inhibitor
الوصف: Background Evidences indicate that inflammatory process plays pivotal role in tumor disease. Soluble epoxide hydrolase inhibitors (sEHIs) have been shown to participate in anti-inflammation and tumorigenesis by protecting epoxyeicosatrienoic acids (EETs). Although we have previously revealed some effects of t-AUCB on glioma in vitro, further investigations are needed to demonstrate its effects on glioblastoma growth in vivo and how to strengthen its antitumor effect. Methods CCK-8 kit was used to test cell growth. Cell migration capacity was performed by wound healing assays. Transwell assay was used to test cell invasion potency. Cell-cycle analysis and cell apoptosis was performed by flow cytometry. The activity of caspase-3 in cells was measured using caspase-3 activity assay kits. Total RNA was extracted from cells lysated by TRIzol reagent. qRT-PCR was performed by ABI 7500 fast RT- PCR system. Lipofectamine RNAiMAX Transfection Reagent (Invitrogen) was used for siRNA transfection. Western blootting was used to test protein expression. Tumor cell xenograft mouse models were used for in vivo study. The SPSS version 17.0 software was applied for statistical analysis. Results Our data shown that t-AUCB inhibits cell proliferation, migration and invasion and induces cell cycle G1 phase arrest in vitro but induces no cell apoptosis; increased Hsp27 activation and following COX-2 overexpression confer resistance to t-AUCB treatment in glioblastoma both in vitro and in vivo; quercetin sensitizes glioblastoma to t-AUCB by dual inhibition of Hsp27 and COX-2 in vitro and in vivo. Conclusions These results indicate that combination of t-AUCB and quercetin may be a potential approach to treating glioblastoma.
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10تقرير
المؤلفون: Deng, Yunyan, Hu, Zhangxi, Shang, Lixia, Chai, Zhaoyang, Tang, Ying Zhong
مصطلحات موضوعية: Scrippsiella trochoidea, resting cyst, dinoflagellate, harmful algal blooms, heat shock protein 20 (Hsp20), heat shock protein 40 (Hsp40), temperature stress, Life Sciences & Biomedicine - Other Topics, Biology
الوصف: The small heat shock protein (sHsp) and Hsp40 are Hsp members that have not been intensively investigated but are functionally important in most organisms. In this study, the potential roles of a Hsp20 (StHsp20) and a Hsp40 (StHsp40) in dinoflagellates during adaptation to temperature fluctuation and alteration of different life stages were explored using the representative harmful algal blooms (HABs)-causative dinoflagellate species, Scrippsiella trochoidea. We isolated the full-length cDNAs of the two genes via rapid amplification of cDNA ends (RACE) and tracked their differential transcriptions via real-time qPCR. The results revealed StHsp20 and StHsp40 exhibited mRNA accumulation patterns that were highly similar in response to heat stress but completely different toward cold stress, which implies that the mechanisms underlying thermal and cold acclimation in dinoflagellates are regulated by different sets of genes. The StHsp20 was probably related to the heat tolerance of the species, and StHsp40 was closely involved in the adaptation to both higher and lower temperature fluctuations. Furthermore, significantly higher mRNA abundance of StHsp40 was detected in newly formed resting cysts, which might be a response to intrinsic stress stemmed from encystment. This finding also implied StHsp40 might be engaged in resting cyst formation of S. trochoidea. Our findings enriched the knowledge about possible cross-talk of different Hsp members in dinoflagellates and provided clues to further explore the molecular underpinnings underlying resting cyst production and broad temperature tolerance of this group of HABs contributors.
العلاقة: BIOLOGY-BASEL; http://ir.qdio.ac.cn/handle/337002/169282Test; http://ir.qdio.ac.cn/handle/337002/169283Test
الإتاحة: https://doi.org/10.3390/biology9110408Test
http://ir.qdio.ac.cn/handle/337002/169282Test
http://ir.qdio.ac.cn/handle/337002/169283Test
