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81.
Methyl CpG-binding proteins induce large-scale chromatin reorganization during terminal differentiation
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Brero A Easwaran HP Nowak D Grunewald I Cremer T Leonhardt H Cardoso MC 《The Journal of cell biology》2005,169(5):733-743
Pericentric heterochromatin plays an important role in epigenetic gene regulation. We show that pericentric heterochromatin aggregates during myogenic differentiation. This clustering leads to the formation of large chromocenters and correlates with increased levels of the methyl CpG-binding protein MeCP2 and pericentric DNA methylation. Ectopic expression of fluorescently tagged MeCP2 mimicked this effect, causing a dose-dependent clustering of chromocenters in the absence of differentiation. MeCP2-induced rearrangement of heterochromatin occurred throughout interphase, did not depend on the H3K9 histone methylation pathway, and required the methyl CpG-binding domain (MBD) only. Similar to MeCP2, another methyl CpG-binding protein, MBD2, also increased during myogenic differentiation and could induce clustering of pericentric regions, arguing for functional redundancy. This MeCP2- and MBD2-mediated chromatin reorganization may thus represent a molecular link between nuclear genome topology and the epigenetic maintenance of cellular differentiation. 相似文献
82.
Inhibition of tumour necrosis factor (TNF)-alpha with biological molecules has proven an effective treatment for rheumatoid
arthritis, achieving a 20% improvement in American College of Rheumatology score in up to 65% of patients. The main drawback
to these and many other biological treatments has been their expense, which has precluded their widespread application. Biological
molecules could alternatively be delivered by gene therapy as the encoding DNA. We have developed novel plasmid vectors termed
pGTLMIK and pGTTMIK, from which luciferase and a dimeric TNF receptor II (dTNFR) are respectively expressed in a doxycycline
(Dox)-regulated manner. Regulated expression of luciferase from the self-contained plasmid pGTLMIK was examined in vitro in a variety of cell lines and in vivo following intramuscular delivery with electroporation in DBA/1 mice. Dox-regulated expression of luciferase from pGTLMIK
of approximately 1,000-fold was demonstrated in vitro, and efficient regulation was observed in vivo. The vector pGTTMIK encoding dTNFR was delivered by the same route with and without administration of Dox to mice with collagen-induced
arthritis. When pGTTMIK was delivered after the onset of arthritis, progression of the disease in terms of both paw thickness
and clinical score was inhibited when Dox was also administered. Vectors with similar regulation characteristics may be suitable
for clinical application. 相似文献
83.
Wilfried Thuiller Luigi Maiorano Florent Mazel Fran?ois Guilhaumon Gentile Francesco Ficetola Sébastien Lavergne Julien Renaud Cristina Roquet David Mouillot 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2015,370(1662)
Protected areas (PAs) are pivotal tools for biodiversity conservation on the Earth. Europe has had an extensive protection system since Natura 2000 areas were created in parallel with traditional parks and reserves. However, the extent to which this system covers not only taxonomic diversity but also other biodiversity facets, such as evolutionary history and functional diversity, has never been evaluated. Using high-resolution distribution data of all European tetrapods together with dated molecular phylogenies and detailed trait information, we first tested whether the existing European protection system effectively covers all species and in particular, those with the highest evolutionary or functional distinctiveness. We then tested the ability of PAs to protect the entire tetrapod phylogenetic and functional trees of life by mapping species'' target achievements along the internal branches of these two trees. We found that the current system is adequately representative in terms of the evolutionary history of amphibians while it fails for the rest. However, the most functionally distinct species were better represented than they would be under random conservation efforts. These results imply better protection of the tetrapod functional tree of life, which could help to ensure long-term functioning of the ecosystem, potentially at the expense of conserving evolutionary history. 相似文献
84.
M. Cristina del Rincn-Castro Jos Barajas-Huerta Jorge E. Ibarra 《Applied microbiology》1999,65(5):2049-2053
Most strains of the insecticidal bacterium Bacillus thuringiensis have a combination of different protoxins in their parasporal crystals. Some of the combinations clearly interact synergistically, like the toxins present in B. thuringiensis subsp. israelensis. In this paper we describe a novel joint activity of toxins from different strains of B. thuringiensis. In vitro bioassays in which we used pure, trypsin-activated Cry1Ac1 proteins from B. thuringiensis subsp. kurstaki, Cyt1A1 from B. thuringiensis subsp. israelensis, and Trichoplusia ni BTI-Tn5B1-4 cells revealed contrasting susceptibility characteristics. The 50% lethal concentrations (LC50s) were estimated to be 4,967 of Cry1Ac1 per ml of medium and 11.69 ng of Cyt1A1 per ml of medium. When mixtures of these toxins in different proportions were assayed, eight different LC50s were obtained. All of these LC50s were significantly higher than the expected LC50s of the mixtures. In addition, a series of bioassays were performed with late first-instar larvae of the cabbage looper and pure Cry1Ac1 and Cyt1A1 crystals, as well as two different combinations of the two toxins. The estimated mean LC50 of Cry1Ac1 was 2.46 ng/cm2 of diet, while Cyt1A1 crystals exhibited no toxicity, even at very high concentrations. The estimated mean LC50s of Cry1Ac1 crystals were 15.69 and 19.05 ng per cm2 of diet when these crystals were mixed with 100 and 1,000 ng of Cyt1A1 crystals per cm2 of diet, respectively. These results indicate that there is clear antagonism between the two toxins both in vitro and in vivo. Other joint-action analyses corroborated these results. Although this is the second report of antagonism between B. thuringiensis toxins, our evidence is the first evidence of antagonism between toxins from different subspecies of B. thuringiensis (B. thuringiensis subsp. kurstaki and B. thuringiensis subsp. israelensis) detected both in vivo and in vitro. Some possible explanations for this relationship are discussed. 相似文献
85.
Giovanni Spagna Riccardo N. Barbagallo Rosa Palmeri Cristina Restuccia Paolo Giudici 《Enzyme and microbial technology》2002,31(7):1030-1035
Three hundred sixty-one yeast strains (80 of which ascribable to Saccharomyces cerevisiae) were isolated from Sicilian musts and wines with the purpose of looking for β-glucosidase (βG, EC 3.2.1.21) activity. Of these, the AL 41 strain had highest endogenous βG activity and was identified as belonging to the species S. cerevisiae by biochemical and molecular methods. This enzyme was subsequently characterized. It had optimum effect at pH 3.5–4.0, whilst optimum temperature was 20 °C, compatible with typical wine-cellar conditions; it was not inhibited by ethanol, at concentrations of 12–14%, or fructose and glucose. The βG was also characterised in terms of the kinetic parameters Km (2.55 mM) and Vmax (1.71 U mg−1 of protein). Finally, it remained stable for at least 35 days in model solutions of must and wine. 相似文献
86.
Timóteo CG Guilherme M Penas D Folgosa F Tavares P Pereira AS 《The Biochemical journal》2012,445(1):125-133
β?-GPI (β?-glycoprotein I) is a plasma glycoprotein ascribed with an anti-angiogenic function; however, the biological role and molecular basis of its action in cell migration remain unknown. The aim of the present study was to assess the contribution of β?-GPI to HAEC (human aortic endothelial cell) migration and the details of its underlying mechanism. Using wound healing and Boyden chamber assays, we found that β?-GPI inhibited endothelial cell migration, which was restored by its neutralizing antibody. NF-κB (nuclear factor κB) inhibitors and lentiviral siRNA (small interfering RNA) silencing of NF-κB significantly attenuated the inhibitory effect of β?-GPI on cell migration. Moreover, β?-GPI was found to induce IκBα (inhibitor of NF-κB) phosphorylation and translocation of p65 and p50. We further demonstrated that mRNA and protein levels of eNOS [endothelial NO (nitric oxide) synthase] and NO production were all increased by β?-GPI and these effects were remarkably inhibited by NF-κB inhibitors and siRNAs of p65 and p50. Furthermore, β?-GPI-mediated inhibition of cell migration was reversed by eNOS inhibitors and eNOS siRNAs. The findings of the present study provide novel insight into the ability of β?-GPI to inhibit endothelial cell migration predominantly through the NF-κB/eNOS/NO signalling pathway, which indicates a potential direction for clinical therapy in vascular diseases. 相似文献
87.
Mayor F Lucas E Jurado-Pueyo M Garcia-Guerra L Nieto-Vazquez I Vila-Bedmar R Fernández-Veledo S Murga C 《Archives of physiology and biochemistry》2011,117(3):125-130
G protein-coupled receptor kinase 2 (GRK2) is emerging as a key, integrative node in many signalling pathways. Besides its canonical role in the modulation of the signalling mediated by many G protein-coupled receptors (GPCR), this protein can display a very complex network of functional interactions with a variety of signal transduction partners, in a stimulus, cell type, or context-specific way. We review herein recent data showing that GRK2 can regulate insulin-triggered transduction cascades at different levels and that this protein plays a relevant role in insulin resistance and obesity in vivo, what uncovers GRK2 as a potential therapeutic target in the treatment of these disorders. 相似文献
88.
Different enzyme requirements for the synthesis of biodiesel: Novozym 435 and Lipozyme TL IM 总被引:5,自引:0,他引:5
Enzymatic syntheses of biodiesel via alcoholysis of different vegetable oils (sunflower, borage, olive and soybean) have been studied. Loss of lipase activity induced by the nucleophile is greater with methanol than with ethanol, and is greater for Lipozyme TL IM than for Novozym 435. The optimum volume of ethanol depends on the loading of solid biocatalyst and is higher for preparations of Novozym 435 than for Lipozyme TL IM. Maximum rates were obtained with Lipozyme TL IM, for a molar ratio of alcohol to FA residues of 0.33. By contrast, Novozym 435 requires at least a 2:1 ratio. Alcoholysis of the vegetable oils is faster with Lipozyme TL IM than with Novozym 435. Use of a high loading of Novozym 435 (50% w/w) and a large molar excess of ethanol are required to obtain an initial rate similar to that obtained with Lipozyme TL IM at a lower enzyme loading (10% w/w) and an equimolar ratio of ethanol and FA residues. Novozym 435 produces quantitative conversions in only 7h at 25 degrees C, but complete conversions are not obtained with Lipozyme TL IM. Three stage stepwise addition of ethanol yields 84% conversion to ethyl esters for Lipozyme TL IM. Hence use of Novozym 435 is preferred. After nine cycles in a batch reactor Novozym 435 retained 85% of its initial activity. 相似文献
89.
Osteoarthritis (OA) is characterized by cartilage degradation. The chondrocyte is the only cell type present in mature cartilage, and it is important in the control of cartilage integrity. The aim of this study was to analyze, by a proteomic approach, the changes that are characteristic of OA chondrocytes, and to identify new OA-related proteins. Chondrocytes were isolated from the cartilage of ten OA patients undergoing joint replacement and ten donors with no history of joint disease. Whole-cell proteins were resolved by 2-DE and stained with SYPRO Ruby. Protein expression patterns of 2-DE gels from OA and normal chondrocyte proteins were analyzed with PDQuest 7.3.1 software. OA-related proteins were identified by MALDI-TOF or MALDI-TOF/TOF MS. The results were validated for ANXA1, GSTO1, GRP78, and HSP90beta in cells by Western blotting and in tissue cartilage by immunohistochemistry. Results showed an average of 700 protein spots that were present in the 2-DE gels. Compared to normal chondrocytes, 19 protein spots were found to be significantly increased in OA cells (ratio OA:N> or =2.0, p<0.05), whereas nine were decreased in OA chondrocytes (ratio OA:N< or =0.5, p<0.05). Three stress response proteins were increased (HSP90beta, GRP78, and GRP94) and three proteins involved in glycolysis were decreased (enolase, glyceraldehyde 3-phosphate dehydrogenase, and fructose biphosphate aldolase). Functionally, almost all proteins could be classified as proteins involved in cellular metabolism (33%), structure (21%), or protein targeting (21%). 相似文献