A lysine substitute for K+. A460K mutation eliminates K+ dependence in H+-pyrophosphatase of Carboxydothermus hydrogenoformans.

The H(+) proton-translocating inorganic pyrophosphatase (H(+)-PPase) family is composed of two phylogenetically distinct types of enzymes: K(+)-dependent and K(+)-independent. However, to date, the sequence criteria governing this dichotomy have remained unknown. In this study, we describe the heterologous expression and functional characterization of H(+)-PPase from the thermophilic bacterium Carboxydothermus hydrogenoformans. Both PP(i)-hydrolyzing and PP(i)-energized H(+) translocation activities of the recombinant enzyme in Escherichia coli inner membrane vesicles are strictly K(+)-dependent. Here we deduce the K(+) requirement of all available H(+)-PPase sequences based on the K(+) dependence of C. hydrogenoformans H(+)-PPase in conjunction with phylogenetic analyses. Our data reveal that K(+)-independent H(+)-PPases possess conserved Lys and Thr that are absent in K(+)-dependent H(+)-PPases. We further demonstrate that a A460K substitution in C. hydrogenoformans H(+)-PPase is sufficient to confer K(+) independence to both PP(i) hydrolysis and PP(i)-energized H(+) translocation. In contrast, a A463T mutation does not affect the K(+) dependence of H(+)-PPase

Detection of Noble Gas Scintillation Light with Large Area Avalanche Photodiodes (LAAPDs)

Large Area Avalanche Photodiodes (LAAPDs) were used for a series of systematic measurements of the scintillation light in Ar, Kr, and Xe gas. Absolute quantum efficiencies are derived. Values for Xe and Kr are consistent with those given by the manufacturer. For the first time we show that argon scintillation (128 nm) can be detected at a quantum efficiency above 40%. Low-pressure argon gas is shown to emit significant amounts of non-UV radiation. The average energy expenditure for the creation of non-UV photons in argon gas at this pressure is measured to be below 378 eV.Comment: 16 pages, 7 figure

Allosteric control of the RNA polymerase by the elongation factor RfaH

Efficient transcription of long polycistronic operons in bacteria frequently relies on accessory proteins but their molecular mechanisms remain obscure. RfaH is a cellular elongation factor that acts as a polarity suppressor by increasing RNA polymerase (RNAP) processivity. In this work, we provide evidence that RfaH acts by reducing transcriptional pausing at certain positions rather than by accelerating RNAP at all sites. We show that 'fast' RNAP variants are characterized by pause-free RNA chain elongation and are resistant to RfaH action. Similarly, the wild-type RNAP is insensitive to RfaH in the absence of pauses. In contrast, those enzymes that may be prone to falling into a paused state are hypersensitive to RfaH. RfaH inhibits pyrophosphorolysis of the nascent RNA and reduces the apparent Michaelis-Menten constant for nucleotides, suggesting that it stabilizes the post-translocated, active RNAP state. Given that the RfaH-binding site is located 75 A away from the RNAP catalytic center, these results strongly indicate that RfaH acts allosterically. We argue that despite the apparent differences in the nucleic acid targets, the time of recruitment and the binding sites on RNAP, unrelated antiterminators (such as RfaH and lambdaQ) utilize common strategies during both recruitment and anti-pausing modification of the transcription complex

Functional specialization of transcription elongation factors

Elongation factors NusG and RfaH evolved from a common ancestor and utilize the same binding site on RNA polymerase (RNAP) to modulate transcription. However, although NusG associates with RNAP transcribing most Escherichia coli genes, RfaH regulates just a few operons containing ops, a DNA sequence that mediates RfaH recruitment. Here, we describe the mechanism by which this specificity is maintained. We observe that RfaH action is indeed restricted to those several operons that are devoid of NusG in vivo. We also show that RfaH and NusG compete for their effects on transcript elongation and termination in vitro. Our data argue that RfaH recognizes its DNA target even in the presence of NusG. Once recruited, RfaH remains stably associated with RNAP, thereby precluding NusG binding. We envision a pathway by which a specialized regulator has evolved in the background of its ubiquitous paralogue. We propose that RfaH and NusG may have opposite regulatory functions: although NusG appears to function in concert with Rho, RfaH inhibits Rho action and activates the expression of poorly translated, frequently foreign genes

On the Underground Production of High Purity Germanium Detectors

Detectors based on high purity germanium (HPGe) are used in numerous deep underground experiments world-wide aiming at detecting rare events like double beta decay and interactions of dark matter. These detectors require the lowest possible background. A significant part of the background is due to radionuclides produced by cosmic-ray interactions with the germanium crystal. This report gives quantitative data on this activation and discusses the possible solutions. The first solution is to optimise the logistics during the crystal and detector fabrication so that the germanium spends a minimum time above ground. The second solution is to implement one or several (up to 12) production steps underground. The report also makes estimates on the future needs for germanium produced underground and the costs involved.JRC.D.4-Isotope measurement

SOLUTION OF THE COUPLED THERMOMECHANICAL PROBLEM OF HYDRODYNAMICS FOR DESIGNING THE PROPULSION SYSTEM OF MICROSATELLITES

This paper considers the problem of predicting the technical and operating characteristics of the propulsion system of a microsatellite created by microelectronic technology. The coupled thermomechanical problem of hydrodynamics has been solved by the ANSYS CFX package. The microengine design has been optimized, the heating of its case in the operating time of the fuel chamber has been analyzed, and the velocity field distribution in the microengine nozzle has been determined. The values of the mechanical and temperature deformation fields for pure silicon and a silicon–SiO2 composite have been found, and the possibility in principle of operation of the considered device has been shown. The calculation was made on a triangular net

Cosmogenic activation of Germanium and its reduction for low background experiments

Production of $^{60}$Co and $^{68}$Ge from stable isotopes of Germanium by nuclear active component of cosmic rays is a principal background source for a new generation of $^{76}$Ge double beta decay experiments like GERDA and Majorana. The biggest amount of cosmogenic activity is expected to be produced during transportation of either enriched material or already grown crystal. In this letter properties and feasibility of a movable iron shield are discussed. Activation reduction factor of about 10 is predicted by simulations with SHIELD code for a simple cylindrical configuration. It is sufficient for GERDA Phase II background requirements. Possibility of further increase of reduction factor and physical limitations are considered. Importance of activation reduction during Germanium purification and detector manufacturing is emphasized.Comment: 10 pages, 3 tables, 6 figure