Mechanism of Na+/H+ antiporting

Na+/H+ antiporters are central to cellular salt and pH homeostasis. The structure of Escherichia coli NhaA was recently determined, but its mechanisms of transport and pH regulation remain elusive. We performed molecular dynamics simulations of NhaA that, with existing experimental data, enabled us to propose an atomically detailed model of antiporter function. Three conserved aspartates are key to our proposed mechanism: Asp164 (D164) is the Na+-binding site, D163 controls the alternating accessibility of this binding site to the cytoplasm or periplasm, and D133 is crucial for pH regulation. Consistent with experimental stoichiometry, two protons are required to transport a single Na+ ion: D163 protonates to reveal the Na+-binding site to the periplasm, and subsequent protonation of D164 releases Na+. Additional mutagenesis experiments further validated the model.     

Intellectual property. Enhanced: intellectual property landscape of the human genome

The impact of gene patents on downstream research and innovation are unknown, in part because of a lack of empirical data on the extent and nature of gene patenting. In this Policy Forum, the authors show that 20% of human gene DNA sequences are patented and that some genes are patented as many as 20 times. Unsurprisingly, genes associated with health and disease are more patented than the genome at large. The intellectual property rights for some genes can become highly fragmented between many owners, which suggests that downstream innovators may face considerable costs to gain access to gene-oriented technologies.     

Dynamic complex formation during the yeast cell cycle

To analyze the dynamics of protein complexes during the yeast cell cycle, we integrated data on protein interactions and gene expression. The resulting time-dependent interaction network places both periodically and constitutively expressed proteins in a temporal cell cycle context, thereby revealing previously unknown components and modules. We discovered that most complexes consist of both periodically and constitutively expressed subunits, which suggests that the former control complex activity by a mechanism of just-in-time assembly. Consistent with this, we show that additional regulation through targeted degradation and phosphorylation by Cdc28p (Cdk1) specifically affects the periodically expressed proteins.     

Accumulation of trehalose in the thermophilic fungus Chaetomium thermophilum var. coprophilum in response to heat or salt stress

The disaccharide trehalose, known to be an effective protectant against various kinds of stress, was observed to accumulate in the cytosol of Chaetomium thermophilum var. coprophilum during heat stress. Trehalose was apparently neither involved in the defence of C. thermophilum var. coprophilum against high concentrations of sodium chloride nor directly linked to thermophily.In C. thermophilum var. coprophilum three different trehalose hydrolyzing activities were eluted from a mono Q anion exchange column by sodium chloride concentrations of 0.10, 0.15 and 0.24 M, respectively.     

Nitrogen acquisition by pea and barley and the effect of their crop residues on available nitrogen for subsequent crops

Nitrogen acquisition by field pea (Pisum sativum L.) and spring barley (Hordeum vulgare L.) grown on a sandy loam soil and availability of N in three subsequent sequences of a cropping system were studied in an outdoor pot experiment. The effect of crop residues on the N availability was evaluated using ¹⁵N-labelled residues. Field pea fixed 75% of its N requirement and the N₂ fixation almost balanced the N removed with the seeds. The barley crop recovered 80% of the ¹⁵N-labelled fertilizer N supplied and the N in the barley grain corresponded to 80% of the fertilizer N taken up by the crop. The uptake of soil-derived N by a test crop (N catch crop) of white mustard (Sinapis alba L.) grown in the autumn was higher after pea than after barley. The N uptake in the test crop was reduced by 27% and 34% after pea and barley residue incorporation, respectively, probably due to N immobilization. The dry matter production and total N uptake of a spring barley crop following pea or barley, with a period of unplanted soil in the autumn/winter, were significantly higher after pea than after barley. The barley crop following pea and barley recovered 11% of the pea and 8% of the barley residue N. The pea and barley residue N recovered constituted only 2.5% and <1%, respectively, of total N in the N-fertilized barley. The total N uptake in a test crop of mustard grown in the second autumn following pea and barley cultivation was not significantly influenced by pre-precrop and residue treatment. In the short term, the incorporation of crop residues was not important in terms of contributing N to the subsequent crop compared to soil and fertilizer N sources, but residues improved the conservation of soil N in the autumn. In the long-term, crop residues are an important factor in maintaining soil fertility and supplying plant-available N via mineralization.     

Mineralization of carbon and nitrogen from fresh and anaerobically stored sheep manure in soils of different texture

A sandy loam soil was mixed with three different amounts of quartz sand and incubated with (¹⁵NH₄)₂SO₄ (60 g N g^-1 soil) and fresh or anaerobically stored sheep manure (60 g g^-1 soil). The mineralization-immobilization of N and the mineralization of C were studied during 84 days of incubation at 20°C. After 7 days, the amount of unlabelled inorganic N in the manure-treated soils was 6–10 g N g^-1 soil higher than in soils amended with only (¹⁵NH₄)₂SO₄. However, due to immobilization of labelled inorganic N, the resulting net mineralization of N from manure was insignificant or slightly negative in the three soil-sand mixtures (100% soil+0% quartz sand; 50% soil+50% quartz sand; 25% soil+75% quartz sand). After 84 days, the cumulative CO₂ evolution and the net mineralization of N from the fresh manure were highest in the soil-sand mixutre with the lowest clay content (4% clay); 28% fo the manure C and 18% of the manure N were net mineralized. There was no significant difference between the soil-sand mixtures containing 8% and 16% clay, in which 24% of the manure C and -1% to 4% of the manure N were net mineralized. The higher net mineralization of N in the soil-sand mixture with the lowest clay content was probably caused by a higher remineralization of immobilized N in this soil-sand mixture. Anaerobic storage of the manure reduced the CO₂ evolution rates from the manure C in the three soil-sand mixtures during the initial weeks of decomposition. However, there was no effect of storage on net mineralization of N at the end of the incubation period. Hence, there was no apparent relationship between net mineralization of manure N and C.