The bZIP class of eukaryotic transcriptional regulators utilize a distinct structural motif that consists of a leucine zipper that mediates dimerization and an adjacent basic region that directly ...contacts DNA. Although models of the protein-DNA complex have been proposed, the basis of DNA-binding specificity is essentially unknown. By genetically selecting for derivatives of yeast GCN4 that activate transcription from promoters containing mutant binding sites, we isolate an altered-specificity mutant in which the invariant asparagine in the basic region of bZIP proteins (Asn-235) has been changed to tryptophan. Wild-type GCN4 binds the optimal site (ATGACTCAT) with much higher affinity than the mutant site (TTGACTCAA), whereas the Trp-235 protein binds these sites with similar affinity. Moreover, the Trp-235, Ala-235, and Gln-235 derivatives differ from GCN4 in their strong discrimination against GTGACTCAC. These results suggest a direct interaction between Asn-235 and the ±4 position of the DNA target site and are discussed in terms of the scissors-grip and induced-fork models of bZIP proteins.
The steady-state translational activation of the GCN4 mRNA is based upon an increase in the rate of ribosome initiation at the protein coding AUG following translation of the 5' most proximal open ...reading frame located in its untranslated region. Such an increase is effected when the cellular amount of the GCN2 protein kinase is increased or when the function of the GCD1 gene product is defective. Here, we report conditions that result in a dramatic transient increase in the rate of GCN4 protein synthesis, which also requires the prior translation of the 5' most proximal open reading frame but is independent of the GCN2 protein. This activation of GCN4 mRNA translation coincides with a decrease in the rate of total cellular protein synthesis. We also observed low rates of protein synthesis in the gcd1 strain and in strains that overexpress the GCN2 protein kinase. The process in protein synthesis that is affected is formation of 43S preinitiation complexes. These results reveal the existence of a coupling between this process in translational initiation and the mechanism that activates translation of GCN4 mRNA.
The present paper reports results from a comparative analysis of the effectiveness of regional policy schemes in areas of Scotland and Greece. Discretionary schemes are of a lower gross cost per job ...than automatic ones in both countries. Additionality was found to be very high among all schemes in both countries. Results indicate that future policy design should be decentralised and highly adapted to local condition s. In the case of Greece, the dominant top-bottom style of policy making acts as major constraints in the adoption of more decentralised policy design and delivery mechanisms. In the case of Scotland such a task is easier to accomplish.
The PICOSEC Micromegas (MM) detector is a precise timing gaseous detector consisting of a Cherenkov radiator combined with a photocathode and a MM amplifying structure. A 100-channel non-resistive ...PICOSEC MM prototype with 10x10 cm^2 active area equipped with a Cesium Iodide (CsI) photocathode demonstrated a time resolution below 18 ps. The objective of this work is to improve the PICOSEC MM detector robustness aspects; i.e. integration of resistive MM and carbon-based photocathodes; while maintaining good time resolution. The PICOSEC MM prototypes have been tested in laboratory conditions and successfully characterised with 150 GeV/c muon beams at the CERN SPS H4 beam line. The excellent timing performance below 20 ps for an individual pad obtained with the 10x10 cm^2 area resistive PICOSEC MM of 20 MOhm/sq showed no significant time resolution degradation as a result of adding a resistive layer. A single-pad prototype equipped with a 12 nm thick Boron Carbide (B4C) photocathode presented a time resolution below 35 ps; opening up new possibilities for detectors with robust photocathodes. The results made the concept more suitable for the experiments in need of robust detectors with good time resolution.
The PICOSEC Micromegas precise timing detector is based on a Cherenkov radiator coupled to a semi-transparent photocathode and a Micromegas amplification structure. The first proof of concept ...single-channel small area prototype was able to achieve time resolution below 25 ps. One of the crucial aspects in the development of the precise timing gaseous detectors applicable in high-energy physics experiments is a modular design that enables large area coverage. The first 19-channel multi-pad prototype with an active area of approximately 10 cm\(^2\) suffered from degraded timing resolution due to the non-uniformity of the preamplification gap. A new 100 cm\(^2\) detector module with 100 channels based on a rigid hybrid ceramic/FR4 Micromegas board for improved drift gap uniformity was developed. Initial measurements with 80 GeV/c muons showed improvements in timing response over measured pads and a time resolution below 25 ps. More recent measurements with a new thinner drift gap detector module and newly developed RF pulse amplifiers show that the resolution can be enhanced to a level of 17~ps. This work will present the development of the detector from structural simulations, design, and beam test commissioning with a focus on the timing performance of a thinner drift gap detector module in combination with new electronics using an automated timing scan method.
The
Saccharomyces cerevisiae GCN4 gene which encodes the transcription activator Gcn4, is under translational regulation. Derepression of
GCN4 mRNA translation is mediated by the Gcn2 protein kinase ...which phosphorylates the α subunit of eIF-2, upon amino-acid starvation. Here, we report that overexpression of certain
Saccharomyces cerevisiae genes generates intracellular conditions that alleviate the requirement for a functional Gcn2 kinase to induce
GCN4 mRNA translation. Our findings, combined with the fact that Gcn2 kinase is dispensable during the initiation phase of the cellular response to amino-acid limitation, provide the grounds to further elucidate the mechanisms underlying the physiology of this homeostatic response.
The yeast gene GCN4 produces an mRNA that has a long 5′ ‘untranslated’ region containing four small open reading frames (ORFs) preceding the protein coding frame. This configuration suppresses the ...rate by which GCN4 protein is synthesized. However, translational derepression of the GCN4 mRNA occurs when yeast cells are grown under conditions of amino acid limitation. Such translational derepression requires the GCN2 protein kinase and the presence of the 5′ most proximal ORF. In this study we show that a functional coupling between the translation of the first ORF and the amount of the GCN2 protein is responsible for the translational derepression of the GCN4 mRNA. Our evidence suggests that this coupling involves an increase in the ability of 40S ribosomal subunits that have translated the first frame to resume scanning and reinitiate translation at a downstream AUG independently of the base sequence in the intervening region.
We describe a proposal to search for an intrinsic electric dipole moment (EDM) of the proton with a sensitivity of \targetsens, based on the vertical rotation of the polarization of a stored proton ...beam. The New Physics reach is of order \(10^~3\)TeV mass scale. Observation of the proton EDM provides the best probe of CP-violation in the Higgs sector, at a level of sensitivity that may be inaccessible to electron-EDM experiments. The improvement in the sensitivity to \(\theta_{QCD}\), a parameter crucial in axion and axion dark matter physics, is about three orders of magnitude.
Results will be shown from the ASTRONEU array developed and operated in the outskirts of Patras, Greece. An array of 9 scintillator detectors and 3 antennas were deployed to study Extensive Air ...Showers (EAS) as a tool for calibrating an underwater neutrino telescope, possible other applications in muon tomography, education purposes, and last but not least, the detection of air showers via their electromagnetic signature. This is the first stage of a total of 24 scintillator counters and 6 RF antennas to complete the array. In this work, results with regard to the electromagnetic detection of showers will be shown. The method of operation and analysis will be presented. The purpose of this project was to demonstrate the adequacy of the method to detect cosmic events even in the presence of high urban electromagnetic background, using noise filters, timing, signal polarization, and eventual comparison with well understood event reconstruction using the scintillator detectors. The results indicate that cosmic showers were detected and the method can be used for the complete array.
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