Error-prone replication of oxidatively damaged DNA by a high-fidelity DNA polymerase

Aerobic respiration generates reactive oxygen species that can damage guanine residues and lead to the production of 8-oxoguanine (8oxoG), the major mutagenic oxidative lesion in the genome. Oxidative damage is implicated in ageing and cancer, and its prevalence presents a constant challenge to DNA polymerases that ensure accurate transmission of genomic information. When these polymerases encounter 8oxoG, they frequently catalyse misincorporation of adenine in preference to accurate incorporation of cytosine. This results in the propagation of G to T transversions, which are commonly observed somatic mutations associated with human cancers. Here, we present sequential snapshots of a high-fidelity DNA polymerase during both accurate and mutagenic replication of 8oxoG. Comparison of these crystal structures reveals that 8oxoG induces an inversion of the mismatch recognition mechanisms that normally proofread DNA, such that the 8oxoG.adenine mismatch mimics a cognate base pair whereas the 8oxoG.cytosine base pair behaves as a mismatch. These studies reveal a fundamental mechanism of error-prone replication and show how 8oxoG, and DNA lesions in general, can form mismatches that evade polymerase error-detection mechanisms, potentially leading to the stable incorporation of lethal mutations.     

Pre-ovipositional and ovipositional behaviour of Lasioseius ometes (Oudemans) and Hypoaspis kargi Costa (Acari: Dermanyssiae: Ascidae,Laelapidae) with notes on egg protection strategies in Mesostigmata

Egg deposition by the soil mites Lasioseius ometes (Oudemans, 1903) and Hypoaspis kargi Costa, 1968 was observed using continuous video-monitoring. The process in both species was found to consist of four distinct phases. In Phase 1, both species engage in pre-ovipositional activity prior to the egg being extruded from the genital opening. In L. ometes, the female often selects a substrate depression for egg laying and examines the depression carefully with the gnathosoma and legs I before oviposition. In H. kargi, egg deposition sites are chosen without regard for substrate depressions. Phase 1 lasted approximately 3 minutes for both species. Phase 2 involved egg extrusion and holding of the egg beneath the gnathosoma. Phase 2 lasted approximately 20 times longer in L. ometes (5 minutes) than in H. kargi. Alteration of the egg surface in the form of thorn-like outgrowths was recorded in L. ometes. Egg deposition occurs in Phase 3, and in L. ometes the egg is generally placed at the deposition site using the chelicerae. Hypoaspis kargi also uses the chelicerae for egg manipulation, but also adjusts orientation of the egg with legs I. The sticky nature of egg surface in H. kargi may provide a means for the female to effectively cover the exposed egg with a protective layer of soil particles. Egg deposition in both species occurred in less than 90 seconds. Phase 4 involves egg covering, but behaviour of the female during this last phase differs markedly between the two species. In L. ometes, the female tends to remain close to the egg, whereas in H. kargi the female moves away from the egg to search for substrate particles appropriate for its covering. The average time for Phase 4 in H. kargi was approximately four times longer (80+ minutes) than in L. ometes.     

Toward a Buddhist Systems Methodology 3: An Application in a Taiwanese Non-Governmental Organization

This paper describes the application of a Buddhist systems methodology (BSM) to tackle a significant conflict (and underlying issues) threatening the future of a large non-governmental Buddhist membership organization in Taiwan. An evaluation of the BSM, undertaken six months after the intervention, demonstrated positive impacts, including a major reduction in conflict; improved communications across the organization (especially from the bottom-up); a successful restructuring to address some of the underlying issues; a significant upturn in the recruitment and retention of members; and a consequential turn-around of the organization's financial position. In addition, several senior managers took on the BSM for their personal use, trained others, and cascaded the methodology down the organization. This resulted in the official adoption of the BSM as the ‘main decision-making system’ for part of the organization, and the start of wider dissemination. Based on these results, the authors argue that the BSM may have more general utility for problem solving and problem prevention in Taiwanese (and possibly other) Buddhist organizations.

Southern Ocean dust-climate coupling over the past four million years

Dust has the potential to modify global climate by influencing the radiative balance of the atmosphere and by supplying iron and other essential limiting micronutrients to the ocean. Indeed, dust supply to the Southern Ocean increases during ice ages, and 'iron fertilization' of the subantarctic zone may have contributed up to 40?parts per million by volume (p.p.m.v.) of the decrease (80-100 p.p.m.v.) in atmospheric carbon dioxide observed during late Pleistocene glacial cycles. So far, however, the magnitude of Southern Ocean dust deposition in earlier times and its role in the development and evolution of Pleistocene glacial cycles have remained unclear. Here we report a high-resolution record of dust and iron supply to the Southern Ocean over the past four million years, derived from the analysis of marine sediments from ODP Site 1090, located in the Atlantic sector of the subantarctic zone. The close correspondence of our dust and iron deposition records with Antarctic ice core reconstructions of dust flux covering the past 800,000 years (refs 8, 9) indicates that both of these archives record large-scale deposition changes that should apply to most of the Southern Ocean, validating previous interpretations of the ice core data. The extension of the record beyond the interval covered by the Antarctic ice cores reveals that, in contrast to the relatively gradual intensification of glacial cycles over the past three million years, Southern Ocean dust and iron flux rose sharply at the Mid-Pleistocene climatic transition around 1.25 million years ago. This finding complements previous observations over late Pleistocene glacial cycles, providing new evidence of a tight connection between high dust input to the Southern Ocean and the emergence of the deep glaciations that characterize the past one million years of Earth history.     

Inclusive fitness theory and eusociality

Arising from M. A. Nowak, C. E. Tarnita & E. O. Wilson 466, 1057-1062 (2010); Nowak et al. reply. Nowak et al. argue that inclusive fitness theory has been of little value in explaining the natural world, and that it has led to negligible progress in explaining the evolution of eusociality. However, we believe that their arguments are based upon a misunderstanding of evolutionary theory and a misrepresentation of the empirical literature. We will focus our comments on three general issues.     

UCP2 mediates ghrelin's action on NPY/AgRP neurons by lowering free radicals

The gut-derived hormone ghrelin exerts its effect on the brain by regulating neuronal activity. Ghrelin-induced feeding behaviour is controlled by arcuate nucleus neurons that co-express neuropeptide Y and agouti-related protein (NPY/AgRP neurons). However, the intracellular mechanisms triggered by ghrelin to alter NPY/AgRP neuronal activity are poorly understood. Here we show that ghrelin initiates robust changes in hypothalamic mitochondrial respiration in mice that are dependent on uncoupling protein 2 (UCP2). Activation of this mitochondrial mechanism is critical for ghrelin-induced mitochondrial proliferation and electric activation of NPY/AgRP neurons, for ghrelin-triggered synaptic plasticity of pro-opiomelanocortin-expressing neurons, and for ghrelin-induced food intake. The UCP2-dependent action of ghrelin on NPY/AgRP neurons is driven by a hypothalamic fatty acid oxidation pathway involving AMPK, CPT1 and free radicals that are scavenged by UCP2. These results reveal a signalling modality connecting mitochondria-mediated effects of G-protein-coupled receptors on neuronal function and associated behaviour.