Phosphorylation networks regulating JNK activity in diverse genetic backgrounds

Cellular signaling networks have evolved to enable swift and accurate responses, even in the face of genetic or environmental perturbation. Thus, genetic screens may not identify all the genes that regulate different biological processes. Moreover, although classical screening approaches have succeeded in providing parts lists of the essential components of signaling networks, they typically do not provide much insight into the hierarchical and functional relations that exist among these components. We describe a high-throughput screen in which we used RNA interference to systematically inhibit two genes simultaneously in 17,724 combinations to identify regulators of Drosophila JUN NH(2)-terminal kinase (JNK). Using both genetic and phosphoproteomics data, we then implemented an integrative network algorithm to construct a JNK phosphorylation network, which provides structural and mechanistic insights into the systems architecture of JNK signaling.     

Drosophila RNAi screen reveals CD36 family member required for mycobacterial infection

Certain pathogens, such as Mycobacterium tuberculosis, survive within the hostile intracellular environment of a macrophage. To identify host factors required for mycobacterial entry and survival within macrophages, we performed a genomewide RNA interference screen in Drosophila macrophage-like cells, using Mycobacterium fortuitum. We identified factors required for general phagocytosis, as well as those needed specifically for mycobacterial infection. One specific factor, Peste (Pes), is a CD36 family member required for uptake of mycobacteria, but not Escherichia coli or Staphylococcus aureus. Moreover, mammalian class B scavenger receptors (SRs) conferred uptake of bacteria into nonphagocytic cells, with SR-BI and SR-BII uniquely mediating uptake of M. fortuitum, which suggests a conserved role for class B SRs in pattern recognition and innate immunity.     

Quantitative morphological signatures define local signaling networks regulating cell morphology

Although classical genetic and biochemical approaches have identified hundreds of proteins that function in the dynamic remodeling of cell shape in response to upstream signals, there is currently little systems-level understanding of the organization and composition of signaling networks that regulate cell morphology. We have developed quantitative morphological profiling methods to systematically investigate the role of individual genes in the regulation of cell morphology in a fast, robust, and cost-efficient manner. We analyzed a compendium of quantitative morphological signatures and described the existence of local signaling networks that act to regulate cell protrusion, adhesion, and tension.     

New surprises in the largest magnetosphere of our solar system

En route to its ultimate rendezvous with Pluto, the New Horizons spacecraft passed through the magnetic and plasma environment of Jupiter in February 2007. Onboard instruments collected high-resolution images, spectroscopic data, and information about charged particles. The results have revealed unusual structure and variation in Jupiter's plasma and large plasmoids that travel down the magnetotail. Data on Jupiter's aurora provide details of the interaction with the solar wind, and a major volcanic eruption from the moon Io was observed during the encounter.     

The FERONIA receptor-like kinase mediates male-female interactions during pollen tube reception

In flowering plants, signaling between the male pollen tube and the synergid cells of the female gametophyte is required for fertilization. In the Arabidopsis thaliana mutant feronia (fer), fertilization is impaired; the pollen tube fails to arrest and thus continues to grow inside the female gametophyte. FER encodes a synergid-expressed, plasma membrane-localized receptor-like kinase. We found that the FER protein accumulates asymmetrically in the synergid membrane at the filiform apparatus. Interspecific crosses using pollen from Arabidopsis lyrata and Cardamine flexuosa on A. thaliana stigmas resulted in a fer-like phenotype that correlates with sequence divergence in the extracellular domain of FER. Our findings show that the female control of pollen tube reception is based on a FER-dependent signaling pathway, which may play a role in reproductive isolation barriers.     

Effects of nitrogen and intensive mixing on decomposition of C-labelled maize (Zea mays L.) residue in soils of different land use types

This study investigated the effects of mineral-N addition and intensive mixing (analogous to disturbance by plowing) on decomposition of ¹⁴C-labelled maize (Zea mays L.) residue and soil organic matter (SOM). Soils were collected from the upper 5 cm of three land use types at Edelweiler, Germany: plow tillage (PT), reduced tillage (RT), and grassland (GL). Soils were incubated for 112 days at 20 °C, with or without ¹⁴C-labelled maize residue (4 g DM kg⁻¹ soil), with or without nitrogen (100 mg N kg⁻¹ soil as NH₄NO₃) and with or without intensive mixing.

The effect of mineral-N on maize residue decomposition differed depending on the stage of decomposition and land use type. Nitrogen accelerated residue decomposition rates in the first 5 days in RT and GL soils, but not in PT soil, and decreased residue decomposition rate in all three land use types after 11 days. At the end of the incubation, N suppressed ¹⁴CO₂ efflux in RT and PT soils, but not in GL soil. Mineral-N did not increase SOM decomposition independently on the land use types.

Intensive mixing stimulated decomposition of both plant residue and SOM in all three land use types. However, effects were smaller in GL soil than in RT or PT soil, presumably because stronger soil aggregates in GL would have been less affected by mixing and allowed greater protection of SOM and plant residue against decomposition.     

A method for determining Michaelis-Menten kinetic parameters of nutrient uptake for plants growing in soil

The relationship between nutrient influx (I_n) and solution concentration at the root surface (C_lo) has not yet been determined for roots growing in soil because of difficulties in measuring C_lo. Corn was grown on two soils with 12 and 21% clay. Each soil had five K levels ranging from low to very high. Potassium influx (I_n) was determined from K uptake between two harvests and root length. C_lo was then calculated from the average soil solution concentration and I_n by assuming that diffusion is the main transport mechanism for K to the root. Potassium influx plotted against C_lo showed a saturation curve, typical of a Michaelis-Menten kinetic relationship. The Michaelis-Menten uptake parameters, maximum influx (I_max) and Michaelis constant (K_m), were obtained by the “Hanes” plot. There was close agreement, without lack of fit, between calculated and observed data. The proposed procedure therefore appears to be suitable for estimating the uptake kinetics of roots growing in soil. Requirements for applications of the method are discussed.     

Accumulation of metals in macrophytes from water reservoirs of a power supply plant, Rio de Janeiro State, Brazil

Aquatic macrophytes are well known accumulators for heavy metals, the reason why they are used as bioindicators for water quality and in phytoremediaton strategies. This study reports on the elemental concentrations in four free-floating aquatic macrophytes (S. auriculata; P. stratiotes; E. crassipes and E. azurea) growing in two water reservoirs (Santana e Vigário, Rio de Janeiro State, Brazil) of an electric power plant that receive input from the polluted Paraíba do Sul River. Filtered water samples and water suspended solids from these environments were also analysed. Inductively coupled plasma mass spectrometry was used as the principal method, allowing the determination of up to 41 elements, including the rare-earth elements (REEs) and other trace metals not assayed before in these macrophytes. The results show that all elements studied are accumulated by the macrophytes with concentration ratios (CR = [plant]: [water]) varying from about 1,000 to 200,000, based on the dry weight of the plant species. With a few exceptions, highest accumulations were observed in E. crassipes in which CRs increase in the sequence: Cu < Mo < Cr < Pb < Tl < Fe < La < Zn < Ce< Mn. Surprisingly high CRs (e.g. Ce: 74,000) and corresponding mass concentrations were observed for the rare-earth elements (e.g. ∑REE: 112 mg kg^?1), also measured in the water suspended particle fraction. The results show that this fraction acts as an effective sink for trace metals in the aquatic system studied and seems to play also an important role in the transfer of metals from water to the plant species.     

Characterization of model melanoidins by the thermal degradation profile

Different types of model melanoidins were thermally degraded, with subsequent identification of the volatiles produced, to obtain and compare the thermal degradation profile of various melanoidins. At first, the volatiles produced from heated glucose/glycine standard melanoidins were compared with glucose/glutamic acid and L-(+)-ascorbic acid/glycine standard melanoidins. In the headspace of heated glucose/glycine melanoidins, mainly furans, were detected, accompanied by carbonyl compounds, pyrroles, pyrazines, pyridines, and some oxazoles. Heating of L-(+)-ascorbic acid/glycine melanoidins resulted in relatively more N-heterocycles, while from glucose/glutamic acid melanoidins no N-heterocycles were formed. In a second part, a chemical treatment was applied to glucose/glycine melanoidins prior to the thermal degradation. Acid hydrolysis was performed to cleave glycosidically linked sugar moieties from the melanoidin skeleton. Nonsoluble glucose/glycine melanoidins were also subjected to an oxidation. The results indicate that the thermal degradation profile is a useful tool in the characterization of different types of melanoidins.     

Untersuchung zur Anwendbarkeit von Sorptionskoeffizienten für die Simulation der Verlagerung von 2, 4, 5-T und LAS in Böden

Investigations for the use of sorption coefficients for simulation of the transport of 2, 4, 5-T and LAS in soils The use of sorption coefficients for the prediction of the transport of 2, 4, 5-trichlorophenoxy-acetic acid (2, 4, 5-T) and of n-dodecylbenzene-sulfonate (LAS) was investigated for four different soil types with the simulation model EXSOL. Linear equilibrium sorption coefficients of 2, 4, 5-T determined in batch studies are not suited for the prediction of the transport wheras values derived from the retention in soil columns better fit the experimental profiles of 2, 4, 5-T in the field. Sorption coefficients for LAS from batch studies with a soil-solution ration of 1:2 and a shaking period of 4 h gave the best results for prediction of LAS transport in the field.