918 resultados para cooperative manipulators


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DNA conformational changes are essential for the assembly of multiprotein complexes that contact several DNA sequence elements. An approach based on atomic force microscopy was chosen to visualize specific protein-DNA interactions occurring on eukaryotic class II nuclear gene promoters. Here we report that binding of the transcription regulatory protein Jun to linearized plasmid DNA containing the consensus AP-1 binding site upstream of a class II gene promoter leads to bending of the DNA template. This binding of Jun was found to be essential for the formation of preinitiation complexes (PICs). The cooperative binding of Jun and PIC led to looping of DNA at the protein binding sites. These loops were not seen in the absence of either PICs, Jun, or the AP-1 binding site, suggesting a direct interaction between DNA-bound Jun homodimers and proteins bound to the core promoter. This direct visualization of functional transcriptional complexes confirms the theoretical predictions for the mode of gene regulation by trans-activating proteins.

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To identify proteins that regulate the transcriptional activity of c-Jun, we have used the yeast two-hybrid screen to detect mammalian polypeptides that might interact functionally with the N-terminal segment of c-Jun, a known regulatory region. Among the proteins identified is a short form of Stat3 (designated Stat3 beta). Stat3 beta is missing the 55 C-terminal amino acid residues of the long form (Stat3 alpha) and has 7 additional amino acid residues at its C terminus. In the absence of added cytokines, expression of Stat3 beta (but not Stat3 alpha) in transfected cells activated a promoter containing the interleukin 6 responsive element of the rat alpha 2-macroglobulin gene; coexpression of Stat3 beta and c-Jun led to enhanced cooperative activation of the promoter. Nuclear extracts of cells transfected with a Stat3 beta expression plasmid formed a complex with an oligonucleotide containing a Stat3 binding site, whereas extracts of cells transfected with a Stat3 alpha plasmid did not. We conclude that there is a short form of Stat3 (Stat3 beta), that Stat3 beta is transcriptionally active under conditions where Stat3 alpha is not, and that Stat3 beta and c-Jun are capable of cooperative activation of certain promoters.

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The Hox gene products are DNA-binding proteins, containing a homeodomain, which function as a class of master control proteins establishing the body plan in organisms as diverse as Drosophila and vertebrates. Hox proteins have recently been shown to bind cooperatively to DNA with another class of homeodomain proteins that include extradenticle, Pbx1, and Pbx2. Hox gene products contain a highly conserved hexapeptide connected by a linker of variable length to the homeodomain. We show that the hexapeptide and the linker region are required for cooperativity with Pbx1 and Pbx2 proteins. Many of the conserved residues present in the Hoxb-8 hexapeptide are required to modulate the DNA binding of the Pbx proteins. Position of the hexapeptide relative to the homeodomain is important. Although deletions of two and four residues of the linker peptide still show cooperative DNA binding, removal of all six linker residues strongly reduces cooperativity. In addition, an insertion of 10 residues within the linker peptide significantly lowers cooperative DNA binding. These results show that the hexapeptide and the position of the hexapeptide relative to the homeodomain are important determinants to allow cooperative DNA binding involving Hox and Pbx gene products.

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The bacteriophage lambda repressor binds cooperatively to pairs of adjacent sites in the lambda chromosome, one repressor dimer binding to each site. The repressor's amino domain (that which mediates DNA binding) is connected to its carboxyl domain (that which mediates dimerization and the interaction between dimers) by a protease-sensitive linker region. We have generated a variant lambda repressor that lacks this linker region. We show that dimers of the variant protein are deficient in cooperative binding to sites at certain, but not all, distances. The linker region thus extends the range over which carboxyl domains of DNA-bound dimers can interact. In particular, the linker is required for cooperative binding to a pair of sites as found in the lambda chromosome, and thus is essential for the repressor's physiological function.

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We have prepared a family of peptide fragments of the 64-residue chymotrypsin inhibitor 2, corresponding to its progressive elongation from the N terminus. The growing polypeptide chain has little tendency to form stable structure until it is largely synthesized, and what structures are formed are nonnative and lack, in particular, the native secondary structural elements of alpha-helix and beta-sheet. These elements then develop as sufficient tertiary interactions are made in the nearly full-length chain. The growth of structure in the small module is highly cooperative and does not result from the hierarchical accretion of substructures.

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The extractive industry, more than any other sector of the economy, often finds itself mired in conflicts with various environmental and community interests. As traditional legal avenues of resolution gave way to the collaborative ideas of alternative dispute resolution, the outcomes, especially the relational outcomes, were less than desirable. This capstone project proposes that an Anticipatory Cooperative Effort (ACE) can help to bridge the gap between industry and environmental interests by encouraging a pro-active and pre-emptive engagement. The point of the ACE concept is not that it defines a new set of principles so much as it repositions where established ADR principles are entertained.

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Traditional visual servoing systems do not deal with the topic of moving objects tracking. When these systems are employed to track a moving object, depending on the object velocity, visual features can go out of the image, causing the fail of the tracking task. This occurs specially when the object and the robot are both stopped and then the object starts the movement. In this work, we have employed a retina camera based on Address Event Representation (AER) in order to use events as input in the visual servoing system. The events launched by the camera indicate a pixel movement. Event visual information is processed only at the moment it occurs, reducing the response time of visual servoing systems when they are used to track moving objects.

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The hexahydride complex OsH6(PiPr3)2 (1) activates the C–OMe bond of 1-(2-methoxy-2-oxoethyl)-3-methylimidazolium chloride (2), in addition to promoting the direct metalation of the imidazolium group, to afford a five-coordinate OsCl(acyl-NHC)(PiPr3)2 (3) compound. The latter coordinates carbon monoxide, oxygen, and molecular hydrogen to give the corresponding carbonyl (4), dioxygen (5), and dihydrogen (6) derivatives. Complex 3 also promotes the heterolytic bond activation of pinacolborane (HBpin), using the acyl oxygen atom as a pendant Lewis base. The hydride ligand and the Bpin substituent of the Fischer-type carbene of the resulting complex 7 activate the O–H bond of alcohols and water. As a consequence, complex 3 is a metal ligand cooperating catalyst for the generation of molecular hydrogen, by means of both the alcoholysis and hydrolysis of pinacolborane, via the intermediates 7 and 6.

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Cooperative learning has been successfully implemented in the last 60 years for teaching at different educational levels including the Higher Studies due to its solid theoretical foundation, the principles it proposes and its practical applications. The purpose of this article is to offer a proposal for some cooperative activities that allow students to work in small groups in a language subject in order to learn not only contents but also putting into practice what they learn, i.e., they learn by being active. This article discusses how the said activities make it possible for students to work with the main principles of cooperative learning, i.e.: positive interdependence, face-to-face interaction, individual and group accountability, interpersonal and small-group skills and group processing. Moreover, this research will also point out that the proposed activities allow students to acquire some of the social competences required in the labour market such as leadership, conflict solving and cooperation.

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Cooperative learning allows students acquisition of competences that are essential for the labour market such as leadership, critical thinking, communication, and so on. For this reason, different cooperative activities were designed in a language subject in English Studies so that students could work in groups and acquire those competences. This article describes some such activities and the emotional competences that students acquire with them. Moreover, a survey was conducted in order to establish students’ opinions about the main competences they acquired with the activities designed and their opinion about a cooperative methodology. Students’ answers were positive and they were aware of what they had learned.