CoA protects against the deleterious effects of caloric overload in Drosophila

We developed a Drosophila model of T2D in which high sugar (HS) feeding leads to insulin resistance. In this model, adipose TG storage is protective against fatty acid toxicity and diabetes. Initial biochemical and gene expression studies suggested that deficiency in CoA might underlie reduced TG synthesis in animals during chronic HS feeding. Focusing on the Drosophila fat body (FB), which is specialized for TG storage and lipolysis, we undertook a series of experiments to test the hypothesis that CoA could protect against the deleterious effects of caloric overload. Quantitative metabolomics revealed a reduction in substrate availability for CoA synthesis in the face of an HS diet. Further reducing CoA synthetic capacity by expressing FB-specific RNAi targeting pantothenate kinase (PK orfumble) or phosphopantothenoylcysteine synthase (PPCS) exacerbated HS-diet-induced accumulation of FFAs. Dietary supplementation with pantothenic acid (vitamin B5, a precursor of CoA) was able to ameliorate HS-diet-induced FFA accumulation and hyperglycemia while increasing TG synthesis. Taken together, our data support a model where free CoA is required to support fatty acid esterification and to protect against the toxicity of HS diets

E2F8 is a nonreceptor activator of heterotrimeric G proteins

BACKGROUND: Heterotrimeric G proteins are important for numerous signaling events in eukaryotes, serving primarily to transduce signals that are initiated by G protein-coupled receptors. It has recently become clear that nonreceptor activators can regulate the level of heterotrimeric G protein signaling and, in some cases, drive cycles of receptor-independent G protein activation. In this study, we used a yeast expression cloning strategy to identify novel nonreceptor activators of heterotrimeric G proteins in a human adipocyte cDNA library. RESULTS: The human transcription factor E2F8 was found to activate heterotrimeric G proteins, suggesting a specific biological role for this recently described member of the E2F family. Epistasis studies showed that E2F8 acted at the level of G proteins and was specific for Gα(i )over Gpa1. E2F8 augmented receptor-driven signaling, but also activated G proteins in the absence of a receptor. The GTPase-activating protein RGS4 antagonized the effect of E2F8, showing that E2F8's effect on Gα involved nucleotide turnover. The entire E2F8 protein was required for full activity, but the majority of the signaling activity appeared to reside in the first 200 residues. CONCLUSION: In yeast, E2F8 is a guanine nucleotide exchange factor (GEF) for the α subunit of heterotrimeric G proteins. The molecular mechanism and biological significance of this effect remain to be determined

Similar effects of high-fructose and high-glucose feeding in a Drosophila model of obesity and diabetes

As in mammals, high-sucrose diets lead to obesity and insulin resistance in the model organism Drosophila melanogaster (called Drosophila hereafter). To explore the relative contributions of glucose and fructose, sucrose’s component monosaccharides, we compared their effects on larval physiology. Both sugars exhibited similar effects to sucrose, leading to obesity and hyperglycemia. There were no striking differences resulting from larvae fed high glucose versus high fructose. Some small but statistically significant differences in weight and gene expression were observed that suggest Drosophila is a promising model system for understanding monosaccharide-specific effects on metabolic homeostasis.</div

Effects of cultivation year and growing location on the phenolic profile of differently coloured carrot cultivars

Carrots (Daucus carota L.) are economically and nutritionally important crops that, apart from carotenoids, contain numerous phenolic compounds which are assumed to exert health beneficial effects. The total phenolic contents of fruits and vegetables are known to depend on cultivar and growing conditions; however, studies examining the variability of a collection of carrots comprising differently coloured cultivars are rare. Therefore, the objective of the present study was to investigate the phenolic compounds of ten differently coloured carrot cultivars considering the effects of three cultivation years at two growing locations. Although total phenolic contents varied in a wide range, both purple cultivars ‘Anthonina’ and ‘Deep Purple’ significantly exceeded those of yellow, orange, red, and uncoloured cultivars (P ≤ 0.05) with amounts from 4,113 to 11,737 mg [kg dry matter (DM)]-1. In contrast to the purple roots, the other generally were characterised by far lower polyphenol contents ranging from 33 to 1,369 mg (kg DM)-1. Interestingly, the values did not considerably vary within these cultivars. In the present study, contrary to cultivar specific effects, the infl uence of growing location was found to be rather weak, supposedly due to similar climatic conditions at both locations. Similarly, variation of phenolic contents from year-to-year was less pronounced. In conclusion, the selection of breeding material was found to be of utmost importance regarding the expression of polyphenols in differently coloured carrots

Diversity, genetic mapping, and signatures of domestication in the carrot (Daucus carota L.) genome, as revealed by Diversity Arrays Technology (DArT) markers

Carrot is one of the most economically important vegetables worldwide, but genetic and genomic resources supporting carrot breeding remain limited. We developed a Diversity Arrays Technology (DArT) platform for wild and cultivated carrot and used it to investigate genetic diversity and to develop a saturated genetic linkage map of carrot. We analyzed a set of 900 DArT markers in a collection of plant materials comprising 94 cultivated and 65 wild carrot accessions. The accessions were attributed to three separate groups: wild, Eastern cultivated and Western cultivated. Twenty-seven markers showing signatures for selection were identified. They showed a directional shift in frequency from the wild to the cultivated, likely reflecting diversifying selection imposed in the course of domestication. A genetic linkage map constructed using 188 F2 plants comprised 431 markers with an average distance of 1.1 cM, divided into nine linkage groups. Using previously anchored single nucleotide polymorphisms, the linkage groups were physically attributed to the nine carrot chromosomes. A cluster of markers mapping to chromosome 8 showed significant segregation distortion. Two of the 27 DArT markers with signatures for selection were segregating in the mapping population and were localized on chromosomes 2 and 6. Chromosome 2 was previously shown to carry the Vrn1 gene governing the biennial growth habit essential for cultivated carrot. The results reported here provide background for further research on the history of carrot domestication and identify genomic regions potentially important for modern carrot breeding