Rotaviruses infect cells by binding to specific cell surface molecules including gangliosides, heat shock protein cognate protein 70 (Hsc70), and some integrins. The characterization of cell surface receptors defining viral tropism is crucial for inhibiting entry into the normal cells or the cancer cells. In the present work, several tumor cell-adapted rotavirus isolates were tested for their interaction with some heat shock proteins (HSPs) present in the U-937 cells, derived from a human pleural effusion (histiocytic lymphoma monocyte). This interaction was examined by virus overlay protein-binding (VOPB), immunochemistry, immuno-dot blot assays, and flow cytometry. The results indicated that the rotavirus isolates studied were able to infect U937 cells by interacting with Hsp90, Hsp70, Hsp60, Hsp40, Hsc70, protein disulfide isomerase (PDI), and integrin β3, which are implicated in cellular proliferation, differentiation, and cancer development. Interestingly, these cellular proteins were found to be associated in lipid microdomains (rafts), facilitating in this way eventual sequential interactions of the rotavirus particles with the cell surface receptors. The rotavirus tropism for U937 cells through the use of these cell surface proteins made this rotavirus isolates an attractive target for the development of oncolytic strategies in the context of alternative and complementary treatment of cancer.
- MeSH
- internalizace viru * MeSH
- lidé MeSH
- membránové proteiny metabolismus MeSH
- nádorové buněčné linie MeSH
- proteiny tepelného šoku HSC70 MeSH
- proteiny tepelného šoku HSP70 genetika MeSH
- proteiny tepelného šoku * metabolismus MeSH
- Rotavirus * metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: The Hsp70 proteins maintain proteome integrity through the capacity of their nucleotide- and substrate-binding domains (NBD and SBD) to allosterically regulate substrate affinity in a nucleotide-dependent manner. Crystallographic studies showed that Hsp70 allostery relies on formation of contacts between ATP-bound NBD and an interdomain linker, accompanied by SBD subdomains docking onto distinct sites of the NBD leading to substrate release. However, the mechanics of ATP-induced SBD subdomains detachment is largely unknown. METHODS: Here, we investigated the structural and allosteric properties of human HSPA1A using hydrogen/deuterium exchange mass spectrometry, ATPase assays, surface plasmon resonance and fluorescence polarization-based substrate binding assays. RESULTS: Analysis of HSPA1A proteins bearing mutations at the interface of SBD subdomains close to the interdomain linker (amino acids L399, L510, I515, and D529) revealed that this region forms a folding unit stabilizing the structure of both SBD subdomains in the nucleotide-free state. The introduced mutations modulate HSPA1A allostery as they localize to the NBD-SBD interfaces in the ATP-bound protein. CONCLUSIONS: These findings show that residues forming the hydrophobic structural unit stabilizing the SBD structure are relocated during ATP-activated detachment of the SBD subdomains to different NBD-SBD docking interfaces enabling HSPA1A allostery. GENERAL SIGNIFICANCE: Mutation-induced perturbations tuned HSPA1A sensitivity to peptide/protein substrates and to Hsp40 in a way that is common for other Hsp70 proteins. Our results provide an insight into structural rearrangements in the SBD of Hsp70 proteins and highlight HSPA1A-specific allostery features, which is a prerequisite for selective targeting in Hsp-related pathologies.
- MeSH
- adenosintrifosfát chemie genetika MeSH
- alosterická regulace genetika MeSH
- konformace proteinů * MeSH
- lidé MeSH
- mutace genetika MeSH
- proteinové domény genetika MeSH
- proteiny tepelného šoku HSP70 chemie genetika MeSH
- vazba proteinů genetika MeSH
- vazebná místa genetika MeSH
- vodík-deuteriová výměna MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cells have elaborated a complex strategy to maintain protein homeostasis under physiological as well as stress conditions with the aim to ensure the smooth functioning of vital processes and producing healthy offspring. Impairment of one of the most important processes in living cells, translation, might have serious consequences including various brain disorders in humans. Here, we describe a variant of the translation initiation factor eIF3a, Rpg1-3, mutated in its PCI domain that displays an attenuated translation efficiency and formation of reversible assemblies at physiological growth conditions. Rpg1-3-GFP assemblies are not sequestered within mother cells only as usual for misfolded-protein aggregates and are freely transmitted from the mother cell into the bud although they are of non-amyloid nature. Their bud-directed transmission and the active movement within the cell area depend on the intact actin cytoskeleton and the related molecular motor Myo2. Mutations in the Rpg1-3 protein render not only eIF3a but, more importantly, also the eIF3 core complex prone to aggregation that is potentiated by the limited availability of Hsp70 and Hsp40 chaperones. Our results open the way to understand mechanisms yeast cells employ to cope with malfunction and aggregation of essential proteins and their complexes.
- MeSH
- eukaryotický iniciační faktor 3 genetika MeSH
- lidé MeSH
- mikrofilamenta genetika MeSH
- mitochondrie MeSH
- mutace MeSH
- myosin typu V genetika MeSH
- proteinové agregáty genetika MeSH
- proteiny tepelného šoku HSP40 genetika MeSH
- proteiny tepelného šoku HSP70 genetika MeSH
- Saccharomyces cerevisiae - proteiny genetika MeSH
- Saccharomyces cerevisiae genetika růst a vývoj MeSH
- těžké řetězce myosinu genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
In the present study, the cDNA sequences of Hsp70 and Hsp90 genes of Isaria farinosa (designated IFHSP70 and IFHSP90) were cloned and characterized using multiple techniques of molecular biology and bioinformatics. The genetic differentiation of the two genes was investigated among 10 geographically separated populations distributed in the Yunnan province. The complete sequence of the IFHSP70 cDNA had a length of 2158 bp, and contained an open reading frame (ORF) of 1962 bp, encoding a 71-kDa polypeptide comprising of 653 amino acids. IFHSP90 cDNA had a length of 2144 bp, and contained an ORF of 2103 bp, encoding a polypeptide of 79.23 kDa, comprising of 700 amino acids. The deduced amino acid sequences of IFHSP70 and IFHSP90 shared high sequence identities with other fungi. Fundamental information pertaining to the protein families, signatures, and conserved motifs of Hsp70 and Hsp90 were also identified. Analysis of molecular variances (AMOVA) from the Hsp70 and Hsp90 genes showed that the genetic variation within-population (83.26%, 83.08%) was greater than among the populations (16.74%, 16.92%). The values of nucleotide diversity (Pi), haplotype diversity (Hd), coefficient of genetic differentiation (Fst), and gene flow (Nm) were calculated. For Hsp70, Pi = 0.0425, Hd = 0.888, Fst = 0.167, Nm = 1.24; For Hsp90, Pi = 0.0420, Hd = 0.894, Fst = 0.169, and Nm = 1.22. These data indicated that the genetic differentiation among 10 different geographical populations of I. farinosa was limited. This study describes, for the first time, cloning, characterization and identification of Isaria farinosa Hsp70 and Hsp90 genes, and provides a preliminary basis for exploring the genetic structure of the genus Isaria using the sequences of Hsp70 and Hsp90 as molecular markers.
- MeSH
- Cordyceps klasifikace genetika metabolismus MeSH
- fungální proteiny chemie genetika metabolismus MeSH
- fylogeneze MeSH
- genetická variace * MeSH
- otevřené čtecí rámce MeSH
- proteiny tepelného šoku HSP70 chemie genetika metabolismus MeSH
- proteiny tepelného šoku HSP90 chemie genetika metabolismus MeSH
- sekvence aminokyselin MeSH
- sekvence nukleotidů MeSH
- sekvenční seřazení MeSH
- výpočetní biologie MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Čína MeSH
Cryptosporidium parvum VF383 has been reported in humans, domesticated ruminants, and wild rats worldwide and described under several names including Cryptosporidium suis-like, based on its close phylogenetic relationship to C. suis. Unlike C. suis, however, it has never been detected in pigs. In the present work, C. parvum VF383 originating from wild brown rats was not infectious for piglets or calves but was infectious for laboratory brown rats, BALB/c mice, and Mongolian gerbils. The prepatent period was 4-5 days for all rodents. The patent period was longer for rats (>30 days) than other rodents (<20 days). None of the rodents developed clinical signs of infection. In all rodents, life cycle stages were detected in the colon by histology and electron microscopy. Oocysts were morphometrically similar to those of C. parvum and smaller than those of C. suis, measuring 5.20 × 4.94 μm. Phylogenetic analyses of 18S rRNA, actin, and HSP70 gene sequences revealed C. parvum VF383 to be genetically distinct from, C. suis, and other described species of Cryptosporidium. Morphological, genetic, and biological data support the establishment of C. parvum VF383 as a new species, and we propose the name Cryptosporidium occultus sp. n.
- MeSH
- aktiny genetika MeSH
- Cryptosporidium klasifikace genetika MeSH
- druhová specificita MeSH
- kolon parazitologie MeSH
- kryptosporidióza parazitologie patologie MeSH
- krysa rodu rattus MeSH
- proteiny tepelného šoku HSP70 genetika MeSH
- RNA ribozomální 18S genetika MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
OBJECTIVES: To analyse the expression regulation of two inducible HSP70 genes - HSPA1A and HSPA1B - located within the major histocompatibility complex (MHC) in patients with various systemic autoimmune diseases and to prove the reliability of MHC-located HSP70 genes as molecular markers reflecting the autoimmune process. METHODS: 94 adult patients with idiopathic inflammatory myopathy (IIM, n=31), systemic lupus erythematosus (SLE, n=31) or systemic sclerosis (SSc, n=32) and 37 healthy individuals were analysed. The mRNA expression level was determined using quantitative real-time PCR method. The expression of intracellular HSP70 was established by flow cytometry, the extracellular HSP70 protein was measured in plasma samples using a commercially available sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: The expression of HSPA1A gene was significantly up-regulated in patients with autoimmune diseases (SLE: p<0.01; SSc: p<0.01; IIM: p<0.0001) compared to healthy controls. The expression of HSPA1B gene was increased only in patients with myositis (p<0.05). Furthermore, the HSPA1B gene expression is associated with the HLA-DRB1*03 risk allele in patients with IIM. In addition, we have found a relation between HSPA1A gene expression regulation and the presence of disease specific autoantibodies in patients with SLE and myositis. The level of intracellular HSP70 was not increased; however, the level of extracellular HSP70 protein was increased in patients suffering from SSc and IIM as compared to controls. CONCLUSIONS: The results suggest an involvement of the MHC-linked HSP70 genes in the pathology of studied autoimmune disorders. Therefore, the HSPA1A and HSPA1B genes might serve as an interesting candidate molecule for development of distinct types of autoimmunities.
- MeSH
- alely MeSH
- autoimunita genetika MeSH
- autoprotilátky MeSH
- biologické markery MeSH
- dospělí MeSH
- genetická predispozice k nemoci MeSH
- lidé středního věku MeSH
- lidé MeSH
- myozitida genetika imunologie MeSH
- proteiny tepelného šoku HSP70 genetika MeSH
- regulace genové exprese MeSH
- senioři MeSH
- systémová sklerodermie genetika imunologie MeSH
- systémový lupus erythematodes genetika imunologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Anaplasma phagocytophilum transmembrane and surface proteins play a role during infection and multiplication in host neutrophils and tick vector cells. Recently,A. phagocytophilumMajor surface protein 4 (MSP4) and Heat shock protein 70 (HSP70) were shown to be localized on the bacterial membrane, with a possible role during pathogen infection in ticks. In this study, we hypothesized thatA. phagocytophilumMSP4 and HSP70 have similar functions in tick-pathogen and host-pathogen interactions. To address this hypothesis, herein we characterized the role of these bacterial proteins in interaction and infection of vertebrate host cells. The results showed thatA. phagocytophilumMSP4 and HSP70 are involved in host-pathogen interactions, with a role for HSP70 during pathogen infection. The analysis of the potential protective capacity of MSP4 and MSP4-HSP70 antigens in immunized sheep showed that MSP4-HSP70 was only partially protective against pathogen infection. This limited protection may be associated with several factors, including the recognition of non-protective epitopes by IgG in immunized lambs. Nevertheless, these antigens may be combined with other candidate protective antigens for the development of vaccines for the control of human and animal granulocytic anaplasmosis. Focusing on the characterization of host protective immune mechanisms and protein-protein interactions at the host-pathogen interface may lead to the discovery and design of new effective protective antigens.
- MeSH
- Anaplasma phagocytophilum genetika metabolismus MeSH
- bakteriální proteiny genetika metabolismus MeSH
- ehrlichióza mikrobiologie veterinární MeSH
- interakce hostitele a patogenu MeSH
- membránové proteiny genetika metabolismus MeSH
- nemoci ovcí mikrobiologie MeSH
- ovce MeSH
- proteiny tepelného šoku HSP70 genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
PROBLEM: Investigating the stress response in the central cotyledon zone of placental tissue in pregnancies with PPROM, PTB, and at term in labor. METHOD OF STUDY: Gene expression of Hsp27, Hsp60, Hsp70, Hsp90, and HspBP1 was compared between these particular groups. Correlation between variables including Hsp gene expression in placental tissue and the gestational age at delivery, WBC count at admission, and serum levels of CRP at admission in patients with PPROM and PTB was determined. RESULTS: Both PPROM and PTB pregnancies were associated with altered Hsp gene expression profile. While PPROM and PTB always induced upregulation of Hsp27 and Hsp60, downregulation of Hsp70 and HspBP1 was present entirely in patients with PPROM. HspBP1 expression profile was also able to differentiate between PPROM and PTB pregnancies. The highest mRNA levels of Hsp60 and Hsp70 were detected in PTB pregnancies with elevated CRP levels at admission. Some of the examined Hsp displayed increased expression with advancing gestational age in both groups (PPROM: Hsp27, Hsp70, and Hsp90; and PTB: Hsp27). CONCLUSION: Upregulation of Hsp27 is a common phenomenon shared between pregnancies affected with PTB and PPROM. On the other hand, downregulation of Hsp70 and HspBP1 represents a unique feature of PPROM.
- MeSH
- adaptorové proteiny signální transdukční genetika metabolismus MeSH
- amnion patologie MeSH
- chaperon hsp60 genetika metabolismus MeSH
- dospělí MeSH
- lidé MeSH
- mitochondriální proteiny genetika metabolismus MeSH
- placenta fyziologie MeSH
- předčasná porodní činnost genetika imunologie MeSH
- proteiny tepelného šoku HSP27 genetika metabolismus MeSH
- proteiny tepelného šoku HSP70 genetika metabolismus MeSH
- proteiny tepelného šoku HSP90 genetika metabolismus MeSH
- regulace genové exprese MeSH
- samovolný potrat genetika imunologie MeSH
- těhotenství MeSH
- transkriptom MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
The EBMT risk score is an established tool successfully used in the prognosis of survival post-HSCT and is applicable for a range of haematological disorders. One of its main advantages is that score generation involves summation of clinical parameters that are available pretransplant. However, the EBMT risk score is recognized as not being optimal. Previous analyses, involving patients with various diagnoses, have shown that non-HLA gene polymorphisms influence outcome after allogeneic HSCT. This study is novel as it focuses only on patients having acute leukaemia (N = 458) and attempts to demonstrate how non-HLA gene polymorphisms can be added to the EBMT risk score in a Cox regression model to improve prognostic ability for overall survival. The results of the study found that three genetic factors improved EBMT risk score. The presence of MAL (rs8177374) allele T in the patient, absence of glucocorticoid receptor haplotype (consisting of rs6198, rs33389 and rs33388) ACT in the patient and absence of heat-shock protein 70-hom (+2437) (rs2227956) allele C in the patient were associated with decreased survival time. When compared to the EBMT risk score, the scores combining EBMT risk score with the genetic factors had an improved correlation with clinical outcome and better separation of risk groups. A bootstrapping technique, involving repeated testing of a model using multiple validation sets, also revealed that the newly proposed model had improved predictive value when compared to the EBMT risk score alone. Results support the view that non-HLA polymorphisms could be useful for pretransplant clinical assessment and provide evidence that polymorphisms in the recipient genotype may influence incoming donor cells, suppressing the initiation of the graft versus leukaemia effect and reducing survival.
- MeSH
- dospělí MeSH
- genomika MeSH
- genotyp MeSH
- haplotypy genetika MeSH
- homologní transplantace škodlivé účinky MeSH
- leukemie genetika imunologie patologie terapie MeSH
- lidé středního věku MeSH
- lidé MeSH
- nemoc štěpu proti hostiteli imunologie MeSH
- prognóza MeSH
- proteiny tepelného šoku HSP70 genetika MeSH
- rizikové faktory MeSH
- testování histokompatibility MeSH
- transplantace hematopoetických kmenových buněk škodlivé účinky MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Při poruchách nálady a neurodegenerativních chorobách nebyly identifikovány geny s přímým patogenetickým vlivem, ale existuje významný genetický podíl pro vznik jak depresivní poruchy, tak Alzheimerovy choroby. Deprese je rizikový faktor, prodrom a doprovodný symptom Alzheimerovy choroby. Mezi geny spojované s rizikem vzniku depresivní poruchy patří SLC64A pro serotoninový transportér (5-HTT), HTR2A pro serotoninový receptor 2A (5-HT2A), TPH2 pro neuronální tryptofanhydroxylázu 2, MAOA pro monoaminooxidázu A, COMT pro katechol-O-metyltransferázu, BDNF pro mozkový neurotrofní faktor, APOE pro apolipoprotein E (ApoE) a další. Hlavním genetickým rizikovým faktorem Alzheimerovy choroby s pozdním nástupem je výskyt ?4-alely APOE. Předložený text přináší přehled faktů o vybraných zkoumaných genetických parametrech a jejich úloze v etiopatogenezi poruch nálady a neurodegenerativních chorob.
Genes have not been identified in mood disorders and neurodegenerative diseases with a direct pathogenetic effect, but the impact of genetic factors is very important in both disorders. Depression is a risk factor, prodromal and also the accompanying symptom of Alzheimer's disease. Genes associated with depression are SLC64A for serotonin transporter (5-HTT), HTR2A for serotonin receptor 2A (5-HT2A), TPH2 for neuronal tryptophanhydroxylase 2, MAOA for monoamine oxidase A, COMT for catechol-O-metyltransferase, BDNF for brain-derived neurotrophic factor, APOE for apolipoprotein E (ApoE) etc. The main genetic risk factor for the development of Alzheimer's disease with the late onset is ?4-alelle of APOE. This review brings the summary of facts about selected examined genetic parameters and their roles in the etiopathogenesis of affective disorders and neurodegenerative disorders.
- MeSH
- Alzheimerova nemoc * etiologie genetika MeSH
- apolipoproteiny E genetika MeSH
- biologické markery MeSH
- depresivní poruchy * etiologie genetika MeSH
- genetická predispozice k nemoci MeSH
- lidé MeSH
- membránové transportní proteiny pro serotonin genetika MeSH
- mozkový neurotrofický faktor genetika MeSH
- polymorfismus genetický MeSH
- proteiny tepelného šoku HSP70 genetika MeSH
- receptor serotoninový 5-HT2A genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
