A Novel Mechanism of Programmed Cell Death in Bacteria by Toxin–Antitoxin Systems Corrupts Peptidoglycan Synthesis

Most genomes of bacteria contain toxin–antitoxin (TA) systems. These gene systems encode a toxic protein and its cognate antitoxin. Upon antitoxin degradation, the toxin induces cell stasis or death. TA systems have been linked with numerous functions, including growth modulation, genome maintenance, and stress response. Members of the epsilon/zeta TA family are found throughout the genomes of pathogenic bacteria and were shown not only to stabilize resistance plasmids but also to promote virulence. The broad distribution of epsilon/zeta systems implies that zeta toxins utilize a ubiquitous bacteriotoxic mechanism. However, whereas all other TA families known to date poison macromolecules involved in translation or replication, the target of zeta toxins remained inscrutable. We used in vivo techniques such as microscropy and permeability assays to show that pneumococcal zeta toxin PezT impairs cell wall synthesis and triggers autolysis in Escherichia coli. Subsequently, we demonstrated in vitro that zeta toxins in general phosphorylate the ubiquitous peptidoglycan precursor uridine diphosphate-N-acetylglucosamine (UNAG) and that this activity is counteracted by binding of antitoxin. After identification of the product we verified the kinase activity in vivo by analyzing metabolite extracts of cells poisoned by PezT using high pressure liquid chromatograpy (HPLC). We further show that phosphorylated UNAG inhibitis MurA, the enzyme catalyzing the initial step in bacterial peptidoglycan biosynthesis. Additionally, we provide what is to our knowledge the first crystal structure of a zeta toxin bound to its substrate. We show that zeta toxins are novel kinases that poison bacteria through global inhibition of peptidoglycan synthesis. This provides a fundamental understanding of how epsilon/zeta TA systems stabilize mobile genetic elements. Additionally, our results imply a mechanism that connects activity of zeta toxin PezT to virulence of pneumococcal infections. Finally, we discuss how phosphorylated UNAG likely poisons additional pathways of bacterial cell wall synthesis, making it an attractive lead compound for development of new antibiotics

A Sustainable Approach for Reusing TreatedWastewater in Agricultural Irrigation in the West Bank – Palestine

There is a critical lack of sanitation in the West Bank (Palestine).Most domestic sewage is disposed of into unlined cesspits or septic tanks, or directly discharged to the environment without treatment. Water resources in the West Bank are limited, and nearly 70% of the water is used for agricultural irrigation. Reuse of treated (reclaimed) wastewater has great potential to alleviate these problems and improve crop yield, but there are many challenges to implementing wastewater reuse. This paper presents a case study in the West Bank town of Tubas, which currently has no sewage collection system or treatment. This study includes traditional engineering design and will address socio-cultural issues through a detailed survey of public perceptions about reclaimed wastewater and an education plan for the various stakeholders in the town. This approach should lead to a wastewater reuse system that is beneficial to Tubas as well as sustainable

Process-based karst modelling to relate hydrodynamic and hydrochemical characteristics to system properties

More than 30% of Europe's land surface is made up of karst exposures. In some countries, water from karst aquifers constitutes almost half of the drinking water supply. Hydrological simulation models can predict the large-scale impact of future environmental change on hydrological variables. However, the information needed to obtain model parameters is not available everywhere and regionalisation methods have to be applied. The responsive behaviour of hydrological systems can be quantified by individual metrics, so-called system signatures. This study explores their value for distinguishing the dominant processes and properties of five different karst systems in Europe and the Middle East. By defining ten system signatures derived from hydrodynamic and hydrochemical observations, a process-based karst model is applied to the five karst systems. In a stepwise model evaluation strategy, optimum parameters and their sensitivity are identified using automatic calibration and global variance-based sensitivity analysis. System signatures and sensitive parameters serve as proxies for dominant processes, and optimised parameters are used to determine system properties. By sensitivity analysis, the set of system signatures was able to distinguish the karst systems from one another by providing separate information about dominant soil, epikarst, and fast and slow groundwater flow processes. Comparing sensitive parameters to the system signatures revealed that annual discharge can serve as a proxy for the recharge area, that the slopes of the high flow parts of the flow duration curves correlate with the fast flow storage constant, and that the dampening of the isotopic signal of the rain as well as the medium flow parts of the flow duration curves have a non-linear relation to the distribution of groundwater storage constants that represent the variability of groundwater flow dynamics. Our approach enabled us to identify dominant processes of the different systems and provided directions for future large-scale simulation of karst areas to predict the impact of future change on karst water resources