the curious paradox of heat shock proteins as tumor specific antigens

... (2009) ATP-binding domain of heat shock protein 70 is essential for its effects on the inhibition of the release of the second mitochondria-derived activator of caspase and apoptosis in C2C12 ... among the regulation of nucleolin ⁄ C23, the activation of caspase-3 and the induction of apoptosis under the setting of oxidative stress and Hsp70 overexpression More importantly, for the first ... nucleus and cytoplasm of the cell, as well as at the cell surface [10] Nucleolin ⁄ C23 regulates many aspects of DNA and RNA metabolism, such as RNA polymerase I-mediated transcription, the folding,...

Ngày tải lên: 16/02/2014, 09:20

... distribution of the size of the inclusions was noticed Moreover, the percentage of cells presenting inclusions as well as the distribution of the size of the inclusions were dependent on when the analysis ... decreased the size of the inclusions but the effect was less intense (average size of 10 lm reduced to about 4–5 lm) In contrast, the size of the inclusions was not significantly altered by the ... was recorded during the FACS analysis The percentage of decrease of DYm was calculated as the ratio of the percentage of cells with FL2-H fluorescence greater than · 101 in the samples to that...

Ngày tải lên: 19/02/2014, 06:20

... Instead of these, mRNA was used as template for the translation With this system, the effect of the CSPs on the translational level was studied In the absence of CSP the expression rate was approximately ... or of masking of regulatory sequences The described translation inhibition of a non-cold shock mRNA can be combined with the results of [3] They found an increase in the translation of cold shock ... and the plasmid pK7CAT The kit was used for the generation of template RNA for in vitro transcription assays and for analysis of the inuence of CSPs at the transcriptory level In the latter case...

Ngày tải lên: 07/03/2014, 21:20

... tuning the AC heating characteristics of SPNPs to improve the efficiency of HSPs 72 induction c Identification of the induction of HSPs 72 d Investigation of the correlation between the change of ... hyperthermia Experimental set-up for AC heat measurements of ferrofluids Formation of colloidal suspension Schematic diagram of the property of SPNPs fluid Kinds of magnetism Hysteresis loops of ... Fe3O4@PEG ferrofluid The specific loss power (SLP) of all the ferrofluids calculated based on the AC heating characteristics obtained in Fig 7-8 Studies of in-vitro biocompatibility of uncoated...

Ngày tải lên: 09/09/2015, 10:08

... recombinant heat shock proteins in the presence or absence of polymyxin B for another 16 h The NF-κB-Luciferase expression was determined using the Dual Luciferase Reporter Assay (DLR Assay, Promega) ... cross-presentation of ovalbumin (OVA) by DC2.4 to the CD8 T cell hybridoma B3Z, as determined by the increased IL-2 production In the presence of 31 the heat shock proteins, B3Z (specific for the SIINFEKL ... expression has led to dramatic decrease of the ability of DC to cross-present peptides [Binder and Srivastava, 2004] However, other receptors for the uptake of the HSP-peptide complexes such as CD40...

Ngày tải lên: 09/10/2015, 11:06

... 2005 FEBS Small heat shock proteins and disease as does the R116C mutation However, in contrast to R116C aA-crystallin, the R49C variant localizes to the cell nucleus and the cytoplasm, superficially ... clearer, as is the molecular basis of oligomerization The implications of oligomer assembly and disassembly as chaperoning prerequisites are under study, sHSP substrates have been identified, and the ... function of the small heat shock protein 16.5 from Methanococcus jannaschii J Biol Chem 277, 38468– 38475 21 Golenhofen N, Perng MD, Quinlan RA & Drenckhahn D (2004) Comparison of the small heat shock...

Ngày tải lên: 19/02/2014, 18:20

... Brg1 during the G1 phase of the cell cycle and regulates the expression of heat shock proteins J Cell Biochem 98, 1528– 1542 26 Kroeger PE, Sarge KD & Morimoto RI (1993) Mouse heat shock transcription ... and heat- shocked at 39 °C for the indicated times Total RNA prepared from the cells was subjected to RT-PCR analysis The genes targeted by yHSF are classified according to the structure of their ... hHSF4–VP16, without changing the magnitude of the heat shock response (cp3) The mutational analysis of the CRYGA and CRYGC HSEs suggests that the nucleotide sequences and configuration of nGAAn-like units...

Ngày tải lên: 07/03/2014, 00:20

... effectiveness of the chaperone Using this assay we tested the relative activity of the two wheat proteins in suppressing aggregation of MDH As shown in Fig 5, both wheat Fig Wheat sHsps suppress heat- induced ... was performed as above except the mobile phase was 200 mM Na/Pi, 100 mM NaCl, pH 7.3 Firefly luciferase reactivation assays Luc was heat- inactivated at 42 °C in the presence of sHsp as described ... TaHsp16.9C-I, suggesting class II proteins have a distinct mode of oligomer assembly as compared to the class I proteins Using malate dehydrogenase (MDH) as a substrate, TaHsp16.9C-I was shown to be a...

Ngày tải lên: 07/03/2014, 15:20

... role of HSP70 in the inhibition of the release of Smac and apoptosis GAPDH Ratio of HSP70 to GAPDH 14 # 12 # 10 * The role of the ATP-binding domain of HSP70 in the prevention of the release of ... (HSP70WT) in other functional assays, including the inhibition of the activation of caspases-9 and -3 (Fig 6A) after exposure to H2O2 for h, as well as the inhibition of H2O2induced apoptosis as assessed ... were assayed for protease activity of caspases-9 or -3 using caspase fluorescent assay kits, and apoptotic cells were identified by elevated activation of caspases-9 and -3 The experiment was repeated...

Ngày tải lên: 16/03/2014, 01:20

... solution of cesium iodide, and processed using MASSLYNX software (Micromass UK Ltd) The peak series were identified manually A mass was generated based on an algorithm built into the MASSLYNX software, ... 25 °C (B) Thermal aggregation of citrate synthase (CS) at 43 °C in the presence of different concentrations of HspB or HspB(G116A) was recorded at 360 nm The CS assay was performed in the absence ... lines assemblies A transfer of CS and HspH into the void volume, at the expense of the other fractions, was confirmed by SDS-PAGE (Fig 7C) Neither HspH nor CS shifted into the void volume when they...

Ngày tải lên: 23/03/2014, 12:20

... units against time in the presence of increasing ratios of MTB HSP 16.3 to CS With the increase of MTB HSP 16.3, there was an increase of protection against thermal aggregation of CS [d, CS alone; ... of stirring The mixture was removed from the ice bath and 200 lL of mgÆmL)1 DNAse was added with stirring for 30 The sample was then placed in a 50-mL tube and centrifuged at 18 000 g for h The ... measure the aggregation of CS in arbitrary units vs the ratios of MTB HSP 16.3/CS and human aB-crystallin/CS With increased ratios of the molecular chaperone protein to CS, there was increased...

Ngày tải lên: 23/03/2014, 21:21

... and the DNA was purified on a 12% acrylamide gel About 50–100 ng of DNA was used for the next cycle of the SELEX procedure Cloning of the products of selection DNA amplified from the last cycle of ... [29] The strategy consists in measuring the affinity of the factors for a series of oligonucleotides containing an increasing number of the binding motif Immediately following the addition of the ... 9) The sequence of JUL, as well as the sequence of Shvalspz, are very close to the consensus binding sequence determined by the SELEX assay (they both contain the 8-mer core), which explains the...

Ngày tải lên: 24/03/2014, 00:21

... function of the small heat shock proteins in plants J Exp Bot 47, 325–338 Arrigo, A.-P & Landry, J (1994) Expression and Function of the Low-molecular-weight Heat Shock Proteins In The Biology of Heat ... and their apparent molecular mass of 350–480 kDa was consistent with oligomers of 20–30 subunits (Fig 3A,E) Thus, the integrity of the extreme N-terminus is not relevant for the assembly of functional ... in a total volume of 1.0 mL of 50 mM phosphate buffer, pH 6.9 CS aggregation was measured by the increase of absorbance at 360 nm in the absence (r) and in the presence of a-Hsp proteins at a final...

Ngày tải lên: 31/03/2014, 21:21

... ON THE ROLES OF UBIQUITIN IN THE REGULATION OF HEAT SHOCK GENE HSP70B’ EE THIAM HEE, GARY NATIONAL UNIVERSITY OF SINGAPORE 2011 INVESTIGATIONS ON THE ROLES OF UBIQUITIN IN THE REGULATION OF HEAT ... REGULATION OF HEAT SHOCK GENE HSP70B’ EE THIAM HEE, GARY (B SCI (HONS), NUS A THESIS SUBMITTED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY DEPARTMENT OF MICROBIOLOGY YONG LOO LIN SCHOOL OF MEDICINE NATIONAL ... DOCTOR OF PHILOSOPHY DEPARTMENT OF MICROBIOLOGY YONG LOO LIN SCHOOL OF MEDICINE NATIONAL UNIVERSITY OF SINGAPORE 2011 ...

Ngày tải lên: 10/09/2015, 08:27

... et al., 2005) Each of these modifications has their role in the regulation of transcription In this project we investigated the role of ubiquitin in the regulation of the heat shock gene HSP70B’ ... The Role of Ubiquitin in Transcriptional Regulation 40 1.5.4 The Proteolytic Role of the Proteosome in the Regulation of Transcription 42 1.5.4 The Non-Proteolytic Role of the Proteosome in the ... Determining the Existence of Ubiquitinated Species of Hsf1 137 3.13 The Stability of Hsf1 141 3.14 The Presence of Hsf1 at the HSP70B’ Promoter 149 vi Chapter 4: Discussion 158 4.1 Key Findings of the...

Ngày tải lên: 10/09/2015, 08:27

... in the template strand to place the coding DNA in the “catalytic cleft” of the RNA polymerase II complex After the synthesis of the first 30 or so bases, RNA polymerase II is thought to release ... the destruction of proteins answered the key criticisms leveled against the Lysosome hypothesis The next question that needed to be answered was what else was there is the elute fraction of the ... was commonly thought of at the time as the typical form of a protease (Ciechanover et al., 1978) Further purification of the flow through revealed an 8.5kDa, protein that was necessary for the...

Ngày tải lên: 10/09/2015, 08:28

... [4] The helical repeat of the DNA (10.5 bp), the length of the unit (5 bp) and the inverted nature of the repeats, positions all of the GAA units on the same side of the double helix, with the ... structure of the yeast Kluyveromyces HSF DBD complexed with the sequence ggTTCtaGAAcc (containing one set of the nGAAn inverted repeat) has been determined [15] In the complex, the third helix of the ... co-crystals, the protein–protein interface consists of the helix amino-terminus, the turn and the wing [15] Unlike other winged helix-turn-helix proteins, the wing of the HSF does not appear to contact the...

Ngày tải lên: 18/02/2014, 04:20

... migration of the spindle during cytokinesis In this case, the depletion of Hsp90a in the absence of HSF1 affects the mitogen-activated protein kinase pathway This study describes the role of HSF1 as ... the loss of other actors in the migration process, such as members of the doublecortin family Dcx and Dclk [146–148] Conclusions The emerging landscape of the role of HSFs in development is the ... binding was shown to be closely associated with reduced methylation of the histone H3K9, irrespective of the relative location of the HSF4-binding regions and of the transcriptional status of genes...

Ngày tải lên: 18/02/2014, 04:20

... measurement of the PGC, the 10 Time (s) 15 20 Fig FCS and FRAP analyses of OlvasGFP in the PGC FCS was used to measure the movement of OlvasGFP into the cytosol out of the nuage region of the PGC Representative ... of OlvasGFP reveals the dynamic nature of the nuage Schematic diagram of the preparation of PGCs of medaka specimen (A) OlvasGFP was expressed in the medaka PGC at stage 24 and localized in the ... measured outside of the nuage in the cytoplasm of the migrating PGC FCS analysis revealed that Olvas GFP diffused with a diffusion constant of 7.0 lm2ặs)1 (Fig 3A) During the measurement of the...

Ngày tải lên: 18/02/2014, 16:20

... displacement of bound proteins from the singlestranded overhang, chiefly hPOT1, as well as possible uncapping of telomerase from the ends There are likely to be multiple mechanisms involved, some of which ... catalytic subunit of telomerase (hTERT in humans) has reverse transcriptase enzymatic activity and synthesizes TTAGGG repeats on to the end of the 3¢ single-stranded overhang Inhibition of hTERT by ... little evidence of concomitant telomere shortening [11,18,32] This has subsequently been shown to be characteristic of the G-quadruplex ligand class as a whole, and the observations of on-target...

Ngày tải lên: 06/03/2014, 09:22