Identification of Isopentenol Biosynthetic Genes from Bacillus subtilis by a Screening Method Based on Isoprenoid Precursor Toxicity

We have developed a novel method to clone terpene synthase genes. This method relies on the inherent toxicity of the prenyl diphosphate precursors to terpenes, which resulted in a reduced-growth phenotype. When these precursors were consumed by a terpene synthase, normal growth was restored. We have demonstrated that this method is capable of enriching a population of engineered Escherichia coli for those clones that express the sesquiterpene-producing amorphadiene synthase. In addition, we enriched a library of genomic DNA from the isoprene-producing bacterium Bacillus subtilis strain 6051 in E. coli engineered to produce elevated levels of isopentenyl diphosphate and dimethylallyl diphosphate. The selection resulted in the discovery of two genes (yhfR and nudF) whose protein products acted directly on the prenyl diphosphate precursors and produced isopentenol. Expression of nudF in E. coli engineered with the mevalonate-based isopentenyl pyrophosphate biosynthetic pathway resulted in the production of isopentenol.     

The brown adipocyte differentiation pathway in birds: An evolutionary road not taken

Background  

Thermogenic brown adipose tissue has never been described in birds or other non-mammalian vertebrates. Brown adipocytes in mammals are distinguished from the more common white fat adipocytes by having numerous small lipid droplets rather than a single large one, elevated numbers of mitochondria, and mitochondrial expression of the nuclear gene UCP1, the uncoupler of oxidative phosphorylation responsible for non-shivering thermogenesis.     

Lipid transformations in greening and senescing leaf tissue

Analyses were made of chlorophyll a and b and fatty acids (18:3, 18:2, 18:1, 18:0, 16:2, 16:1, and 16:0) of greening and senescing leaf tissue. Those dark-grown tissues given a prior treatment of red, far red, or red followed by far red light showed similar increases in chlorophylls and linolenate (18:3) when exposed to continuous white light. In contrast, green barley (Hordeum vulgare L.) leaves placed in the dark lost chlorophylls and fatty acids, especially 18:3. Senescing cocklebur (Xanthium strumarium L.) leaf tissue showed a decline in chlorophyll and fatty acids, especially again 18:3. Abscisic acid, but not sucrose, accelerated these senescent changes. Radioactive acetate incorporation into the galacto-lipids and phospholipids of senescing cocklebur leaf tissue increased and then the radioactivity of the lipids decreased in senescent tissues.     

The inshore fish faunas over soft substrates and reefs on the tropical west coast of Australia differ and change with latitude and bioregion

Aim To test the following hypotheses regarding ichthyofaunal compositions along an extensive tropical coastline. The compositions over soft substrates and reefs: (1) consistently differ markedly; (2) change progressively with latitude and temperature through sequential changes in the abundances of certain species; and (3) vary among bioregions, as those regions differ markedly in their environmental characteristics. Location Tropical north‐western Australia. Methods Similarity matrices, derived from percentage contributions of each fish species to catches obtained over soft substrates by trawling and over reefs by trapping at seven regularly spaced sites along 1500 km of coast, were subjected to cluster analysis, non‐metric multidimensional scaling ordination and associated tests. Results In total, 361 species were obtained by trawling and trapping along the tropical coast of north‐western Australia (NWA). Only 56 species were recorded over both soft substrates and reefs, whereas 229 and 76 species were caught exclusively over soft substrates and reefs, respectively. The Leiognathidae, Carangidae and Terapontidae contributed most individuals (62.2%) to catches over soft substrates, whereas the Lethrinidae and Lutjanidae dominated those over reefs (81.9%). The species compositions in both habitats were related to latitude and water temperature. Ichthyofaunal compositions in the Kimberley region differed markedly from those in the Canning/Pilbara regions further south, which, in turn, each had distinct characters. The vast majority of species found over both habitats also occur in the Pacific Ocean to the north. Main conclusions The most important fish families over soft substrates and reefs in inshore marine waters of tropical NWA differ markedly. The ichthyofaunal compositions of both habitats undergo similar patterns of progressive change with latitude, due to site‐staggered changes in the relative abundances of key fish species in their respective habitats. Ichthyofaunal composition in both habitats was found to be influenced by water temperature. The latitudinal trends exhibited by species composition are overlaid on a strong bioregional effect, reflecting the influence of the very different environmental conditions in those bioregions, which include marked differences in such factors as tidal regime, turbidity and whether mangroves are nearby. The important contribution of species with a Pacific affinity was presumably facilitated by the polewards‐flowing Indonesian Throughflow, which links the Pacific Ocean with waters bordering the NWA coast.     

Tri-locus sequence data reject a “Gondwanan origin hypothesis” for the African/South Pacific crab genus Hymenosoma

Crabs of the family Hymenosomatidae are common in coastal and shelf regions throughout much of the southern hemisphere. One of the genera in the family, Hymenosoma, is represented in Africa and the South Pacific (Australia and New Zealand). This distribution can be explained either by vicariance (presence of the genus on the Gondwanan supercontinent and divergence following its break-up) or more recent transoceanic dispersal from one region to the other. We tested these hypotheses by reconstructing phylogenetic relationships among the seven presently-accepted species in the genus, as well as examining their placement among other hymenosomatid crabs, using sequence data from two nuclear markers (Adenine Nucleotide Transporter [ANT] exon 2 and 18S rDNA) and three mitochondrial markers (COI, 12S and 16S rDNA). The five southern African representatives of the genus were recovered as a monophyletic lineage, and another southern African species, Neorhynchoplax bovis, was identified as their sister taxon. The two species of Hymenosoma from the South Pacific neither clustered with their African congeners, nor with each other, and should therefore both be placed into different genera. Molecular dating supports a post-Gondwanan origin of the Hymenosomatidae. While long-distance dispersal cannot be ruled out to explain the presence of the family Hymenosomatidae on the former Gondwanan land-masses and beyond, the evolutionary history of the African species of Hymenosoma indicates that a third means of speciation may be important in this group: gradual along-coast dispersal from tropical towards temperate regions, with range expansions into formerly inhospitable habitat during warm climatic phases, followed by adaptation and speciation during subsequent cooler phases.     

Conformational features of crystal-surface cellulose from higher plants

Native cellulose in higher plants forms crystalline fibrils a few nm across, with a substantial fraction of their glucan chains at the surface. The accepted crystal structures feature a flat-ribbon 21 helical chain conformation with every glucose residue locked to the next by hydrogen bonds from O-3' to O-5 and from O-2 to O-6'. Using solid-state NMR spectroscopy we show that the surface chains have a different C-6 conformation so that O-6 is not in the correct position for the hydrogen bond from O-2. We also present evidence consistent with a model in which alternate glucosyl residues are transiently or permanently twisted away from the flat-ribbon conformation of the chain, weakening the O-3' - 0-5 hydrogen bond. Previous molecular modelling and the modelling studies reported here indicate that this 'translational' chain conformation is energetically feasible and does not preclude binding of the surface chains to the interior chains, because the surface chains share the axial repeat distance of the 21 helix. Reduced intramolecular hydrogen bonding allows the surface chains to form more hydrogen bonds to external molecules in textiles, wood, paper and the living plant.     

Regulation of the synthesis of cyclic glucan in Xanthomonas campestris by a diffusible signal molecule

The rpf gene cluster of Xanthomonas campestris pv. campestris is involved in the co-ordinate positive regulation of the production of extracellular enzymes and the extracellular polysaccharide xanthan. Several of the rpf genes are involved in a regulatory system involving the small diffusible molecule DSF (for diffusible signal factor). Synthesis of DSF requires RpfF, and a two-component sensory transduction system involving RpfC has been implicated in the perception of the signal and signal transduction. Here we show that mutations in both rpfF and rpfC lead to reductions in the levels of cyclic glucan. The levels of cyclic glucan synthetase in membrane preparations from rpfF and rpfC mutants were, however, unaltered from the wild-type. Similar alterations in the level of cyclic glucan without changes in cyclic glucan synthetase activity were seen when wild-type bacteria were exposed to osmotic stress. These results extend the range of cellular functions subject to regulation by the rpf genes and DSF system.     

Nutritional challenges during development induce sex-specific changes in glucose homeostasis in the adult sheep

The early-life environment has implications for risk of adult-onset diseases, such as glucose intolerance, insulin insensitivity, and obesity, effects that may occur with or without reduced birth weight. We determined the consequences of nutrient restriction in early gestation and early postnatal life and their interactions on postnatal growth, body composition, and glucose handling. Ewes received 100% (C, n = 39) or 50% nutritional requirements (U, n = 41) from 1 to 31 days gestation and 100% thereafter. Male and female offspring (singleton/twin) from C and U ewes were then fed either ad libitum (CC n = 22, UC n = 19) or to reduce body weight to 85% of target from 12 to 25 wk of age (CU n = 17, UU n = 22) and ad libitum thereafter. At 1.5 and 2.5 yr, glucose handling was determined by area under the curve (AUC) for glucose and insulin concentrations following intravenous glucose (0.5 g/kg body wt). Insulin sensitivity was determined at 2.5 yr following intravenous insulin (0.5 IU/kg). In females, postnatal undernutrition reduced (P < 0.05) glucose AUC at both ages, regardless of prenatal nutrition. Postnatal undernutrition did not affect insulin secretion in females but enhanced insulin-induced glucose disappearance in singletons. Poor early postnatal growth was associated with increased fat in females. In males, glucose tolerance was unaffected by undernutrition despite changes in insulin AUC dependent on age, treatment, and single/twin birth. Nutrition in early postnatal life has long-lasting, sex-specific effects on glucose handling in sheep, likely due, in females, to enhanced insulin sensitivity. Improved glucose utilization may aid weight recovery but have negative implications for glucose homeostasis and body composition over the longer term.     

Lipoprotein(a) in women twins: heritability and relationship to apolipoprotein(a) phenotypes.

Lp(a) is a unique lipoprotein consisting of an LDL-like particle and a characteristic protein, apo(a). Increased levels of Lp(a) constitute a risk factor for coronary heart disease. Variation in the size of the apo(a) protein is a phenotype controlled by the apo(a) gene on chromosome 6 and is related to Lp(a) plasma levels. Based on 169 MZ and 125 DZ adult female twin pairs, this study's purpose was to estimate the proportion of the variation in Lp(a) levels that is due to genetic influences and to determine the extent to which the apo(a) locus explains this heritability. Lp(a) levels were significantly more similar in MZ twins than in DZ twins: mean co-twin differences were 3.9 +/- 5.7 mg/dl and 16.0 +/- 19.9 mg/dl (P less than .001), respectively. Intraclass correlations were .94 in MZ twins and .32 in DZ twins, resulting in a heritability estimate of .94 (P less than .001). Heritability was then calculated using only co-twins with the same apo(a) phenotype: the heritability estimate decreased to .45 but was still highly significant (P less than .001). Therefore, on the basis of heritability analysis of women twins, Lp(a) levels are almost entirely genetically controlled. Variation at the apo(a) locus contributes to this heritability, although other genetic factors could be involved.