A data set of human endogenous protein ubiquitination sites

Lysine ubiquitination is an important and versatile protein post-translational modification. Numerous cellular functions are regulated by ubiquitination, suggesting that extensive numbers of proteins, if not all, are modified with ubiquitin at certain times. However, proteome-wide profiling of ubiquitination sites in the mammalian system is technically challenging. We report the design and characterization of an engineered protein affinity reagent for the isolation of ubiquitinated proteins and the identification of ubiquitination sites with mass spectrometry. This recombinant protein consists of four tandem repeats of ubiquitin-associated domain from UBQLN1 fused to a GST tag. We used this GST-qUBA reagent to isolate polyubiquitinated proteins and identified 294 endogenous ubiquitination sites on 223 proteins from human 293T cells without proteasome inhibitors or overexpression of ubiquitin. Mitochondrial proteins constitute 14.7% of this data set, implicating ubiquitination in a wide range of mitochondrial functions.     

A new approach to determining environmental flow requirements: Sustaining the natural values of floodplains of the southern Murray‐Darling Basin

Summary Large overbank flood events play an important role in maintaining large‐scale ecological processes and connectivity along and across the floodplains and between the rivers and their floodplains in the southern Murray‐Darling Basin. However, the regulation of rivers means that extensive overbank flooding can only occur in the rare circumstance of extreme flood events. Recent environmental water allocations have focussed on the largest floodplain blocks (‘icon’ sites) and a small set of specific values (e.g. colonial nesting waterbirds), as well as on trialling fine‐scale manipulation of infrastructure (e.g. pumping) to water relatively small areas. There has been no comprehensive systematic assessment of the entire floodplain and its wider set of flood‐dependent natural assets (such as ecosystems and species; herein referred to as ‘natural values’) to maximise the effectiveness of environmental water use and to catalogue values likely to be lost. This paper describes an assessment of some 220 000 ha found to support flood‐dependent natural values in Victoria. We mapped the geographic distribution and estimated components of the flooding requirements (natural flooding frequency, and maximum period without flooding and minimum duration of each flooding event before significant deterioration) for each natural value. Using an example of one stretch of the River Murray, we show how the resultant spatial data can be used with floodplain inundation modelling to compare the outcomes of real or planned environmental watering events; potentially providing tools for management agencies to conserve a wider range of floodplain values than is currently the case. That is, water managers and the public can see what ecosystems and threatened species are intended to be maintained by environmental watering and what values are intended to be abandoned across the whole floodplain, rather than just seeing the small subset of values and ‘icon’ sites that are intended to be maintained. Examples are provided to illustrate how information about the location, water requirements and extent covered by potential floods for specific values can be used to build adaptive watering strategies for areas as large as the whole floodplain.     

A global reorganization of the metabolome in Arabidopsis during cold acclimation is revealed by metabolic fingerprinting

Many plants, including Arabidopsis , increase their freezing tolerance in response to low, non-freezing temperatures. This process is known as cold acclimation and involves many complex biochemical changes at the level of the metabolome. Our goal was to examine the effects of cold acclimation on the metabolome using a non-targeted metabolic fingerprinting approach. Multivariate data analyses indicate that, in Arabidopsis, a global reprogramming of metabolism occurs as a result of cold acclimation. By measuring an entire spectrum of putative metabolites based on mass-to-charge ( m / z ) ratios, vs. an individual or group of metabolite(s), a comprehensive, unbiased assessment of metabolic processes relative to cold acclimation was determined. Whereas leaves shifted to low temperature present metabolic profiles that are constantly changing, leaves developed at low temperature demonstrate a stable complement of components. Although it appears that some metabolic networks are modulated by the environment, others require development under low-temperature conditions for adjustment. Understanding how metabolism as a whole is regulated allows the integration of cellular, physiological and ecological attributes in a biological system, a necessity if complex traits, such as freezing tolerance, are to be modified by breeding or genetic manipulation.     

Isosteric ramatroban analogs: selective and potent CRTH-2 antagonists

The chemoattractant receptor-homologous molecule expressed on T(H)2 cells (CRTH-2), also found on eosinophils and basophils, is a prostaglandin D2 receptor involved in the recruitment of these cell types during an inflammatory response. In this report, we describe the synthesis and optimization of a ramatroban isostere that is a selective and potent antagonist of CRTH-2 which may be useful in the treatment of certain diseases.     

Pharmacogenomic profiling of an oxidative stress-mediated spongiform encephalopathy

The majority of cellular superoxide is generated in the mitochondria as a by-product of normal oxidative metabolism. In the mitochondria, superoxide is detoxified by manganese superoxide dismutase (SOD2). Mice lacking SOD2 demonstrate a multifaceted neonatal lethal phenotype, including a spongiform encephalopathy that is preventable through antioxidant treatment. The molecular events behind the observed pathology in the cortex of these mice are unknown. We hypothesized that the lack of SOD2 would result in significant changes in cortical gene expression and that therapeutically beneficial antioxidant treatment would normalize the expression of some genes, providing insight into the mechanism by which mitochondrial oxidative stress results in neurodegeneration. We report the identification of gene expression profiles associated with this paradigm, which characterize the degree of response to the pharmacologic intervention. We have identified specific pathways targeted by endogenous oxidative stress, including glutathione metabolism, iron metabolism, and cell-survival pathways centering on the kinase AKT. The normalization of expression of some of these pathways by antioxidant treatment suggests approaches to treating disease in which endogenous oxidative stress plays a role.     

The Open Microscopy Environment (OME) Data Model and XML file: open tools for informatics and quantitative analysis in biological imaging

The Open Microscopy Environment (OME) defines a data model and a software implementation to serve as an informatics framework for imaging in biological microscopy experiments, including representation of acquisition parameters, annotations and image analysis results. OME is designed to support high-content cell-based screening as well as traditional image analysis applications. The OME Data Model, expressed in Extensible Markup Language (XML) and realized in a traditional database, is both extensible and self-describing, allowing it to meet emerging imaging and analysis needs.     

Diversity within cyanobacterial mat communities in variable salinity meltwater ponds of McMurdo Ice Shelf, Antarctica

This study investigated the diversity of cyanobacterial mat communities of three meltwater ponds--Fresh, Orange and Salt Ponds, south of Bratina Island, McMurdo Ice Shelf, Antarctica. A combined morphological and genetic approach using clone libraries was used to investigate the influence of salinity on cyanobacterial diversity within these ecosystems without prior cultivation or isolation of cyanobacteria. We were able to identify 22 phylotypes belonging to Phormidium sp., Oscillatoria sp. and Lyngbya sp. In addition, we identified Antarctic Nostoc sp., Nodularia sp. and Anabaena sp. from the clone libraries. Fresh (17 phylotypes) and Orange (nine phylotypes) Ponds showed a similar diversity in contrast to that of the hypersaline Salt Pond (five phylotypes), where the diversity within cyanobacterial mats was reduced. Using the comparison of identified phylotypes with existing Antarctic sequence data, it was possible to gain further insight into the different levels of distribution of phylotypes identified in the investigated cyanobacterial mat communities of McMurdo Ice Shelf.     

Ground predator abundance affects prey removal in highbush blueberry (<Emphasis Type="BoldItalic">Vaccinium corymbosum</Emphasis>) fields and can be altered by aisle ground covers

Habitat management to conserve natural enemies has increased biological control of insect pests in various cropping systems [Annu. Rev. Entomol. 45: 175–201, 2000]. We wanted to determine if insect predation in highbush blueberry, Vaccinium corymbosum L. (Ericales: Ericaceae), is influenced by manipulation of edaphic arthropod community and whether management of ground cover in aisles between blueberry rows enhances this community. The first question was studied in blueberry plots bounded by trenches permitting selective movement into plots (ingress) or out of plots (egress), as well as unbounded control plots. We observed a significant effect of boundary type on the arthropod communities relative abundance as measured with pitfall traps, with relative abundance highest in ingress plots, intermediate in control plots and lowest in egress plots. Effects of ground arthropod abundance on predation rates were assessed with onion fly, Delia antiqua (Meigen) (Diptera: Anthomyiidae), pupae as sentinel prey. Pupa recovery was greatest in egress boundary plots, intermediate in control plots and lowest in ingress boundary plots. Regression analyses indicate pupal recovery rate decreased as a function of carabid abundance as well as the abundance of non-insect ground predators. To determine if ground cover management influenced natural enemy abundance, aisles were clean cultivated or planted with three ground covers (clover, ryegrass, or buckwheat). Increasing ground cover had a significant effect on the relative abundance of Harpalus pensylvanicus De Geer (Coleoptera: Carabidae). In addition to conserving natural enemies for control of blueberry insect pests, we discuss additional benefits of ground covers that may increase their utility for blueberry production.