Insulin regulates menin expression, cytoplasmic localization, and interaction with FOXO1

Menin is the ubiquitously expressed nuclear protein product of the MEN1 gene, which interacts with PKB/Akt in the cytoplasm to inhibit its activity. This study describes a novel insulin-dependent mechanism of menin regulation and interaction with other metabolic proteins. We show that insulin downregulated menin in a time-dependent manner via the human insulin receptor. Inhibition analysis indicated a critical role for the protein kinase Akt in regulation of menin expression and localization. Insulin-mediated decrease in menin expression was abrogated by the PI3K/Akt inhibitor LY-294002 at early time points, from 2 to 7 h. Furthermore, exposure to insulin resulted in the cytoplasmic localization of menin and increased interaction with FOXO1. Fasting followed by refeeding modulates serum insulin levels, which corresponded to an increase in menin interaction with FOXO1 in the liver. Liver-specific hemizygous deletion of menin resulted in increased expression of FOXO1 target genes, namely IGFBP-1, PGC-1α, insulin receptor, Akt, and G-6-Pase. This study provides evidence that menin expression and localization are regulated by insulin signaling and that this regulation triggers an increase in its interaction with FOXO1 via Akt with metabolic consequences.     

High-yield production of hydrophobins RodA and RodB from <Emphasis Type="Italic">Aspergillus fumigatus</Emphasis> in <Emphasis Type="Italic">Pichia pastoris</Emphasis>

Hydrophobins are small fungal proteins with amphipatic properties and the ability to self-assemble on a hydrophobic/hydrophilic interface; thus, many technical applications for hydrophobins have been suggested. The pathogenic fungus Aspergillus fumigatus expresses the hydrophobins RodA and RodB on the surface of its conidia. RodA is known to be of importance to the pathogenesis of the fungus, while the biological role of RodB is currently unknown. Here, we report the successful expression of both hydrophobins in Pichia pastoris and present fed-batch fermentation yields of 200–300 mg/l fermentation broth. Protein bands of expected sizes were detected by SDS-PAGE and western blotting, and the identity was further confirmed by tandem mass spectrometry. Both proteins were purified using his-affinity chromatography, and the high level of purity was verified by silver-stained SDS-PAGE. Recombinant RodA as well as rRodB were able to convert a glass surface from hydrophilic to hydrophobic similar to native RodA, but only rRodB was able to decrease the hydrophobicity of a Teflon-like surface to the same extent as native RodA, while rRodA showed this ability to a lesser extent. Recombinant RodA and native RodA showed a similar ability to emulsify air in water, while recombinant RodB could also emulsify oil in water better than the control protein bovine serum albumin (BSA). This is to our knowledge the first successful expression of hydrophobins from A. fumigatus in a eukaryote host, which makes it possible to further characterize both hydrophobins. Furthermore, the expression strategy and fed-batch production using P. pastoris may be transferred to hydrophobins from other species.     

Acquisition of phosphorus and other poorly mobile nutrients by roots. Where do plant nutrition models fail?

Background

In the context of increasing global food demand, ecological intensification of agroecosystems is required to increase nutrient use efficiency in plants while decreasing fertilizer inputs. Better exploration and exploitation of soil resources is a major issue for phosphorus, given that rock phosphate ores are finite resources, which are going to be exhausted in decades from now on.

Scope

We review the processes governing the acquisition by plants of poorly mobile nutrients in soils, with a particular focus on processes at the root?Csoil interface. Rhizosphere processes are poorly accounted for in most plant nutrition models. This lack largely explains why present-day models fail at predicting the actual uptake of poorly mobile nutrients such as phosphorus under low input conditions. A first section is dedicated to biophysical processes and the spatial/temporal development of the rhizosphere. A second section concentrates on biochemical/biogeochemical processes, while a third section addresses biological/ecological processes operating in the rhizosphere.

Conclusions

New routes for improving soil nutrient efficiency are addressed, with a particular focus on breeding and ecological engineering options. Better mimicking natural ecosystems and exploiting plant diversity appears as an appealing way forward, on this long and winding road towards ecological intensification of agroecosystems.     

Beyond species area curves: application of a scale‐free measure for spatial variability of species richness

We report a novel pattern in species richness, complementary to the well‐known species–area relationship. We show that, as sample area increases, the variation in relative richness decreases among otherwise comparable spatial units. This pattern holds for southern African birds, French birds, Cape Proteaceae and the trees of Barro Colorado Island. We propose a scale‐free method for quantifying this pattern by measuring the multifractal intensity of species richness, which is the multi‐scale tendency of adjacent patches with the same area to differ in richness. By this measure, spatial variability is strongest for Cape Proteaceae and weakest for Barro Colorado Island trees. Our results have implications for area‐dependent estimates of species‐richness, for example in reserve planning and in simulation‐based studies. They imply that such estimates are most accurate for large areas, and will be subject to substantial uncertainty when the multifractal intensity is high and the area is small. For comparative purposes, multifractal intensity may be used as a supplement or as an alternative to mean richness, as well as for other ecological densities, such as biomass distribution and local abundance.     

Reproduction of the spiny puffer, Diodon holocanthus (Pisces: Diodontidae) in the continental shelf of Mexican Central Pacific

Diodon holocanthus is an important economic and ecological species of the demersal fish community, caught as bycatch from local shrimp fishery. The reproductive biology of this long-spine porcupinefish has not yet been described, and reproductive season, the sex ratio, length distribution, length at first gonad maturity, and the gonad macro and microscopic features are described. A total of 400 organisms, ranging from 5.0 to 40.3 cm (average 18.4 cm) total length, were caught from the continental shelf of the Central Mexican Pacific, from December 1995 and December 1998. Sex ratio was 1:0.86 females to males (n = 253). The length at which 50% of the individuals showed maturing gonads was 19.7 cm for females and 20.1 cm for males. Length of the smallest organism with ripe gonads was 12.2 cm for females and 13 cm for males. Four gonadal maturation stages were found in both sexes, and five oocyte development phases were identified. The oocyte development pattern is of asynchronous type, which means the species can reproduce several times a year. Testicle development is lobular type, as in most teleost fishes. Monthly mean values of the gonad-somatic index suggest the reproduction activity peaks in June, and September-December.     

Maternal protein restriction elevates cholesterol in adult rat offspring due to repressive changes in histone modifications at the cholesterol 7alpha-hydroxylase promoter

Adverse events in utero, such as intrauterine growth restriction (IUGR), can permanently alter epigenetic mechanisms leading to the metabolic syndrome, which encompasses a variety of symptoms including augmented cholesterol. The major site for cholesterol homeostasis occurs via the actions of hepatic cholesterol 7α-hydroxylase (Cyp7a1), which catabolizes cholesterol to bile acids. To determine whether posttranslational histone modifications influence the long-term expression of Cyp7a1 in IUGR, we used a protein restriction model in rats. This diet during pregnancy and lactation led to IUGR offspring with decreased liver to body weight ratios, followed by increased circulating and hepatic cholesterol levels in both sexes at d 21 and exclusively in the male offspring at d 130. The augmented cholesterol was associated with decreases in the expression of Cyp7a1. Chromatin immunoprecipitation revealed that this was concomitant with diminished acetylation and enhanced methylation of histone H3 lysine 9 [K9,14], markers of chromatin silencing, surrounding the promoter region of Cyp7a1. These epigenetic modifications originate in part due to dietary-induced decreases in fetal hepatic Jmjd2a expression, a histone H3 [K9] demethylase. Collectively, these findings suggest that the augmented cholesterol observed in low-protein diet-derived offspring is due to permanent repressive posttranslational histone modifications at the promoter of Cyp7a1. Moreover, this is the first study to demonstrate that maternal undernutrition leads to long-term cholesterol dysregulation in the offspring via epigenetic mechanisms.