Analysis of jak2 catalytic function by peptide microarrays: The role of the JH2 domain and V617F mutation

Janus kinase 2 (JAK2) initiates signaling from several cytokine receptors and is required for biological responses such as erythropoiesis. JAK2 activity is controlled by regulatory proteins such as Suppressor of Cytokine Signaling (SOCS) proteins and protein tyrosine phosphatases. JAK2 activity is also intrinsically controlled by regulatory domains, where the pseudokinase (JAK homology 2, JH2) domain has been shown to play an essential role. The physiological role of the JH2 domain in the regulation of JAK2 activity was highlighted by the discovery of the acquired missense point mutation V617F in myeloproliferative neoplasms (MPN). Hence, determining the precise role of this domain is critical for understanding disease pathogenesis and design of new treatment modalities. Here, we have evaluated the effect of inter-domain interactions in kinase activity and substrate specificity. By using for the first time purified recombinant JAK2 proteins and a novel peptide micro-array platform, we have determined initial phosphorylation rates and peptide substrate preference for the recombinant kinase domain (JH1) of JAK2, and two constructs comprising both the kinase and pseudokinase domains (JH1-JH2) of JAK2. The data demonstrate that (i) JH2 drastically decreases the activity of the JAK2 JH1 domain, (ii) JH2 increased the Kmfor ATP (iii) JH2 modulates the peptide preference of JAK2 (iv) the V617F mutation partially releases this inhibitory mechanism but does not significantly affect substrate preference or Kmfor ATP. These results provide the biochemical basis for understanding the interaction between the kinase and the pseudokinase domain of JAK2 and identify a novel regulatory role for the JAK2 pseudokinase domain. Additionally, this method can be used to identify new regulatory mechanisms for protein kinases that provide a better platform for designing specific strategies for therapeutic approaches

A magnetic shield/dual purpose mission

The objective of this work is to design, build, and fly a dual-purpose payload whose function is to produce a large volume, low intensity magnetic field and to test the concept of using such a magnetic field to protect manned spacecraft against particle radiation. An additional mission objective is to study the effect of this moving field on upper atmosphere plasmas. Both mission objectives appear to be capable of being tested using the same superconducting coil. The potential benefits of this magnetic shield concept apply directly to both earth-orbital and interplanetary missions. This payload would be a first step in assessing the true potential of large volume magnetic fields in the U.S. space program. Either converted launch systems or piggyback payload opportunities may be appropriate for this mission. The use of superconducting coils for magnetic shielding against solar flare radiation during manned interplanetary missions has long been contemplated and was considered in detail in the years preceding the Apollo mission. With the advent of new superconductors, it has now become realistic to reconsider this concept for a Mars mission. Even in near-earth orbits, large volume magnetic fields produced using conventional metallic superconductors allow novel plasma physics experiments to be contemplated. Both deployed field-coil and non-deployed field-coil shielding arrangements have been investigated, with the latter being most suitable for an initial test payload in a polar orbit

Significant Impacts of Increasing Aridity on the Arid Soil Microbiome

Global deserts occupy one-third of the Earth’s surface and contribute significantly to organic carbon storage, a process at risk in dryland ecosystems that are highly vulnerable to climate-driven ecosystem degradation. The forces controlling desert ecosystem degradation rates are poorly understood, particularly with respect to the relevance of the arid-soil microbiome. Here we document correlations between increasing aridity and soil bacterial and archaeal microbiome composition along arid to hyperarid transects traversing the Atacama Desert, Chile. A meta-analysis reveals that Atacama soil microbiomes exhibit a gradient in composition, are distinct from a broad cross-section of nondesert soils, and yet are similar to three deserts from different continents. Community richness and diversity were significantly positively correlated with soil relative humidity (SoilRH). Phylogenetic composition was strongly correlated with SoilRH, temperature, and electrical conductivity. The strongest and most significant correlations between SoilRH and phylum relative abundance were observed for Acidobacteria, Proteobacteria, Planctomycetes, Verrucomicrobia, and Euryarchaeota (Spearman’s rank correlation [rs] = >0.81; false-discovery rate [q] = ≤0.005), characterized by 10- to 300-fold decreases in the relative abundance of each taxon. In addition, network analysis revealed a deterioration in the density of significant associations between taxa along the arid to hyperarid gradient, a pattern that may compromise the resilience of hyperarid communities because they lack properties associated with communities that are more integrated. In summary, results suggest that arid-soil microbiome stability is sensitive to aridity as demonstrated by decreased community connectivity associated with the transition from the arid class to the hyperarid class and the significant correlations observed between soilRH and both diversity and the relative abundances of key microbial phyla typically dominant in global soils. Importance: We identify key environmental and geochemical factors that shape the arid soil microbiome along aridity and vegetation gradients spanning over 300 km of the Atacama Desert, Chile. Decreasing average soil relative humidity and increasing temperature explain significant reductions in the diversity and connectivity of these desert soil microbial communities and lead to significant reductions in the abundance of key taxa typically associated with fertile soils. This finding is important because it suggests that predicted climate change-driven increases in aridity may compromise the capacity of the arid-soil microbiome to sustain necessary nutrient cycling and carbon sequestration functions as well as vegetative cover in desert ecosystems, which comprise one-third of the terrestrial biomes on Earth.</p

Development of NEXRAD Wind Retrievals as Input to Atmospheric Dispersion Models

The objective of this study is to determine the feasibility that routinely collected data from the Doppler radars can appropriately be used in Atmospheric Dispersion Models (ADMs) for emergency response. We have evaluated the computational efficiency and accuracy of two variational mathematical techniques that derive the u- and v-components of the wind from radial velocities obtained from Doppler radars. A review of the scientific literature indicated that the techniques employ significantly different approaches in applying the variational techniques: 2-D Variational (2DVar), developed by NOAA¹s (National Oceanic and Atmospheric Administration's) National Severe Storms Laboratory (NSSL) and Variational Doppler Radar Analysis System (VDRAS), developed by the National Center for Atmospheric Research (NCAR). We designed a series of numerical experiments in which both models employed the same horizontal domain and resolution encompassing Oklahoma City for a two-week period during the summer of 2003 so that the computed wind retrievals could be fairly compared. Both models ran faster than real-time on a typical single dual-processor computer, indicating that they could be used to generate wind retrievals in near real-time. 2DVar executed ~2.5 times faster than VDRAS because of its simpler approach