Arabidopsis annexin1 mediates the radical-activated plasma membrane Ca2+ - and K+ -permeable conductance in root cells

Plant cell growth and stress signaling require Ca2+ influx through plasma membrane transport proteins that are regulated by reactive oxygen species. In root cell growth, adaptation to salinity stress, and stomatal closure, such proteins operate downstream of the plasma membrane NADPH oxidases that produce extracellular superoxide anion, a reactive oxygen species that is readily converted to extracellular hydrogen peroxide and hydroxyl radicals, OH_. In root cells, extracellular OH_ activates a plasma membrane Ca2+-permeable conductance that permits Ca2+ influx. In Arabidopsis thaliana, distribution of this conductance resembles that of annexin1 (ANN1). Annexins are membrane binding proteins that can form Ca2+-permeable conductances in vitro. Here, the Arabidopsis loss-of-function mutant for annexin1 (Atann1) was found to lack the root hair and epidermal OH_-activated Ca2+- and K+-permeable conductance. This manifests in both impaired root cell growth and ability to elevate root cell cytosolic free Ca2+ in response to OH_. An OH_-activated Ca2+ conductance is reconstituted by recombinant ANN1 in planar lipid bilayers. ANN1 therefore presents as a novel Ca2+-permeable transporter providing a molecular link between reactive oxygen species and cytosolic Ca2+ in plants

The Protein Model Portal

Structural Genomics has been successful in determining the structures of many unique proteins in a high throughput manner. Still, the number of known protein sequences is much larger than the number of experimentally solved protein structures. Homology (or comparative) modeling methods make use of experimental protein structures to build models for evolutionary related proteins. Thereby, experimental structure determination efforts and homology modeling complement each other in the exploration of the protein structure space. One of the challenges in using model information effectively has been to access all models available for a specific protein in heterogeneous formats at different sites using various incompatible accession code systems. Often, structure models for hundreds of proteins can be derived from a given experimentally determined structure, using a variety of established methods. This has been done by all of the PSI centers, and by various independent modeling groups. The goal of the Protein Model Portal (PMP) is to provide a single portal which gives access to the various models that can be leveraged from PSI targets and other experimental protein structures. A single interface allows all existing pre-computed models across these various sites to be queried simultaneously, and provides links to interactive services for template selection, target-template alignment, model building, and quality assessment. The current release of the portal consists of 7.6 million model structures provided by different partner resources (CSMP, JCSG, MCSG, NESG, NYSGXRC, JCMM, ModBase, SWISS-MODEL Repository). The PMP is available at http://www.proteinmodelportal.org and from the PSI Structural Genomics Knowledgebase

Teachers in an Interdisciplinary Learning Community: Engaging, Integrating, and Strengthening K-12 Education

This study examines the inputs (processes and strategies) and outputs (perceptions, skill development, classroom transfer, disciplinary integration, social networking, and community development) of a yearlong, interdisciplinary teacher learning and development experience. Eleven secondary math and science teachers partnered with an interdisciplinary team of university engineering mentors in a yearlong engineering education and project implementation program. It consisted of a 6-week on-site resident professional development and collaboration experience, with an ongoing support and follow-up including digital systems. Mixed-method, multisource data indicate that teachers engaged with motivations combining personal, intrinsic interest and classroom integration goals. They formed and sustained an active community of learning and practice that supported their success, on-site and through classroom integration, thereby promoting innovations. Teachers reported positive perceptions throughout the program and demonstrated significant, productive trajectories of change-over-time. Teachers learned and transferred task-specific engineering and scientific skills, as well as more general inquiry-based pedagogical strategies to their secondary classrooms.Yeshttps://us.sagepub.com/en-us/nam/manuscript-submission-guideline

Towards a New Paradigm for Intuitive Theatrical Lighting Control

A simplified model of a lighting process applied in theatrical productions is one that involves two key players. The first is that of the lighting designer, to produce a set of intentions and plans for the scenes that define the show. The second, the lighting technician, has the job of translating these designs into practice using control equipment, luminaires, and other technical instruments. The lighting design often becomes a ‘working document’ subject to change and adaptation as the physical reality of the design becomes apparent, and the input of other stakeholders is considered. This process can be a valuable creative tool, and also a difficult technical hurdle to overcome, depending on a varied number of factors. A common frustration with this process is that either the complexity of the task, or difficulty in communication can make it difficult for the final creative vision to be effectively realised. Strains may also arise in the case of small, often touring, theatre companies where the lighting designer and technician may be the same person, and frequently one of the performers as well. Considering the design aspect, there can be challenges in ensuring efficacy of lighting plans between venues in touring productions, with 2D lighting sketches or even 3D computer simulations confined to the paper or screen. From a technical perspective, the role of the lighting technician in theatres and performance situations has included the operation of lighting control equipment during shows. The equipment has evolved over time but has, until recently, been grounded upon the basis of faders and the mixing desk. It is argued that this paradigm has failed to keep pace with the change in other interactive technologies. The on-going research described in this paper explores existing and upcoming technologies in the field, whilst also seeking to understand the roles and communication workflows of those involved in theatrical lighting to find the best areas to seek improvement, adopting principles of user-centred design. The intention of this research is to develop a new paradigm, and manifestation of it, using a control method for lighting or projection that allows a more intuitive form of operation in theatre productions, which will be scalable and flexible

Modern Electronic Techniques Applied to Physics and Engineering

Contains reports on seven research projects.Office of Scientific Research and Development (OSRD) OEMsr-26

Genome variation and population structure among 1142 mosquitoes of the African malaria vector species Anopheles gambiae and Anopheles coluzzii

Mosquito control remains a central pillar of efforts to reduce malaria burden in sub-Saharan Africa. However, insecticide resistance is entrenched in malaria vector populations, and countries with a high malaria burden face a daunting challenge to sustain malaria control with a limited set of surveillance and intervention tools. Here we report on the second phase of a project to build an open resource of high-quality data on genome variation among natural populations of the major African malaria vector species Anopheles gambiae and Anopheles coluzzii. We analyzed whole genomes of 1142 individual mosquitoes sampled from the wild in 13 African countries, as well as a further 234 individuals comprising parents and progeny of 11 laboratory crosses. The data resource includes high-confidence single-nucleotide polymorphism (SNP) calls at 57 million variable sites, genome-wide copy number variation (CNV) calls, and haplotypes phased at biallelic SNPs. We use these data to analyze genetic population structure and characterize genetic diversity within and between populations. We illustrate the utility of these data by investigating species differences in isolation by distance, genetic variation within proposed gene drive target sequences, and patterns of resistance to pyrethroid insecticides. This data resource provides a foundation for developing new operational systems for molecular surveillance and for accelerating research and development of new vector control tools. It also provides a unique resource for the study of population genomics and evolutionary biology in eukaryotic species with high levels of genetic diversity under strong anthropogenic evolutionary pressures

Of Bits and Bugs — On the Use of Bioinformatics and a Bacterial Crystal Structure to Solve a Eukaryotic Repeat-Protein Structure

Pur-α is a nucleic acid-binding protein involved in cell cycle control, transcription, and neuronal function. Initially no prediction of the three-dimensional structure of Pur-α was possible. However, recently we solved the X-ray structure of Pur-α from the fruitfly Drosophila melanogaster and showed that it contains a so-called PUR domain. Here we explain how we exploited bioinformatics tools in combination with X-ray structure determination of a bacterial homolog to obtain diffracting crystals and the high-resolution structure of Drosophila Pur-α. First, we used sensitive methods for remote-homology detection to find three repetitive regions in Pur-α. We realized that our lack of understanding how these repeats interact to form a globular domain was a major problem for crystallization and structure determination. With our information on the repeat motifs we then identified a distant bacterial homolog that contains only one repeat. We determined the bacterial crystal structure and found that two of the repeats interact to form a globular domain. Based on this bacterial structure, we calculated a computational model of the eukaryotic protein. The model allowed us to design a crystallizable fragment and to determine the structure of Drosophila Pur-α. Key for success was the fact that single repeats of the bacterial protein self-assembled into a globular domain, instructing us on the number and boundaries of repeats to be included for crystallization trials with the eukaryotic protein. This study demonstrates that the simpler structural domain arrangement of a distant prokaryotic protein can guide the design of eukaryotic crystallization constructs. Since many eukaryotic proteins contain multiple repeats or repeating domains, this approach might be instructive for structural studies of a range of proteins

A Novel Acyl-CoA Beta-Transaminase Characterized from a Metagenome

BACKGROUND: Bacteria are key components in all ecosystems. However, our knowledge of bacterial metabolism is based solely on the study of cultivated organisms which represent just a tiny fraction of microbial diversity. To access new enzymatic reactions and new or alternative pathways, we investigated bacterial metabolism through analyses of uncultivated bacterial consortia. METHODOLOGY/PRINCIPAL FINDINGS: We applied the gene context approach to assembled sequences of the metagenome of the anaerobic digester of a municipal wastewater treatment plant, and identified a new gene which may participate in an alternative pathway of lysine fermentation. CONCLUSIONS: We characterized a novel, unique aminotransferase that acts exclusively on Coenzyme A (CoA) esters, and proposed a variant route for lysine fermentation. Results suggest that most of the lysine fermenting organisms use this new pathway in the digester. Its presence in organisms representative of two distinct bacterial divisions indicate that it may also be present in other organisms