Ubiquitin‐Activated Interaction Traps (UBAITs) identify E3 ligase binding partners

We describe a new class of reagents for identifying substrates, adaptors, and regulators of HECT and RING E3s. UBAITs (Ub iquitin‐A ctivated I nteraction T raps) are E3‐ubiquitin fusion proteins and, in an E1‐ and E2‐dependent manner, the C‐terminal ubiquitin moiety forms an amide linkage to proteins that interact with the E3, enabling covalent co‐purification of the E3 with partner proteins. We designed UBAITs for both HECT (Rsp5, Itch) and RING (Psh1, RNF126, RNF168) E3s. For HECT E3s, trapping of interacting proteins occurred in vitro either through an E3 thioester‐linked lariat intermediate or through an E2 thioester intermediate, and both WT and active‐site mutant UBAITs trapped known interacting proteins in yeast and human cells. Yeast Psh1 and human RNF126 and RNF168 UBAITs also trapped known interacting proteins when expressed in cells. Human RNF168 is a key mediator of ubiquitin signaling that promotes DNA double‐strand break repair. Using the RNF168 UBAIT, we identify H2AZ—a histone protein involved in DNA repair—as a new target of this E3 ligase. These results demonstrate that UBAITs represent powerful tools for profiling a wide range of ubiquitin ligases.     

Bioactivities of simplified adociaquinone B and naphthoquinone derivatives against Cdc25B,MKP-1, and MKP-3 phosphatases

Some simplified adociaquinone B analogs and a series of 1,4-naphthoquinone derivatives were synthesized and tested against the three enzymes Cdc25B, MKP-1, and MKP-3. Cdc25B and MKP-1 in particular are enzymes overexpressed in human cancer cells, and they represent potential molecular targets for novel cancer chemotherapeutic treatments. A number of analogs exhibited significant inhibitory activity against these enzymes, and the bioassay data in addition to structure–activity relationships of these compounds will be discussed.     

Extensive Reorganization of Behavior Accompanies Ontogeny of Aggression in Male Flesh Flies

Aggression, costly in both time and energy, is often expressed by male animals in defense of valuable resources such as food or potential mates. Here we present a new insect model system for the study of aggression, the male flesh fly Sarcophaga crassipalpis, and ask whether there is an ontogeny of aggression that coincides with reproductive maturity. After establishing that reproductive maturity occurs by day 3 of age (post-eclosion), we examined the behavior of socially isolated males from different age cohorts (days 1, 2, 3, 4, and 6) upon introduction, in a test arena, with another male of the same age. The results show a pronounced development of aggression with age. The change from relative indifference to heightened aggression involves a profound increase in the frequency of high-intensity aggressive behaviors between days 1 and 3. Also noteworthy is an abrupt increase in the number of statistically significant transitions involving these full-contact agonistic behaviors on day 2. This elevated activity is trimmed back somewhat by day 3 and appears to maintain a stable plateau thereafter. No convincing evidence was found for escalation of aggression nor the establishment of a dominance relationship over the duration of the encounters. Despite the fact that aggressive interactions are brief, lasting only a few seconds, a major reorganization in the relative proportions of four major non-aggressive behaviors (accounting for at least 96% of the total observation time for each age cohort) accompanies the switch from low to high aggression. A series of control experiments, with single flies in the test arenas, indicates that these changes occur in the absence of the performance of aggressive behaviors. This parallel ontogeny of aggressive and non-aggressive behaviors has implications for understanding how the entire behavioral repertoire may be organized and reorganized to accommodate the needs of the organism.     

Spatial and temporal variation in butterfly biodiversity in a West African forest: lessons for establishing efficient rapid monitoring programmes

Because tropical forests face serious threats and are usually situated in developing countries, cheap and easy Rapid Biodiversity Assessment (RBA) seems an ideal strategy for documenting changes in habitat quality and setting conservation priorities. Fruit‐feeding butterflies are often used as focal taxa to document biodiversity trends in tropical forests. However, the choice of inappropriate sampling strategies may nullify its value. More extensive data sets that sample multiple months and seasons and incorporate the vertical dimension are needed as a baseline for designing reliable sampling regimes. Therefore, we performed a butterfly trapping study in Ghana, using both understorey and canopy traps, and sampling seven periods covering dry and wet seasons. Both individual numbers and species richness were on average three to four times higher in the understorey than in the canopy traps with strikingly different species composition (only 11% overlap in species). The number of species and individual abundance recorded in a month varied extensively. This study underlined the importance of taking into account temporal variation and vertical stratification when designing RBAs of fruit‐feeding butterflies in West African forests. We recommend the use of both understorey and canopy traps and inclusion of both ‘wet’ and ‘dry’ seasons into RBA sampling regimes.     

Elevated Density of [3H]Imipramine Binding in Aged Human Brain

Aging was associated with an increase in the density of specific binding sites for [3H]imipramine in postmortem specimens of human hypothalamus, frontal cortex, and parietal cortex. In general, [3H]imipramine binding was not affected by factors considered difficult to control in postmortem studies, i.e., time from death to autopsy and cause of death. The in vitro regulation of [3H]imipramine binding by sodium was impaired with age in hypothalamic homogenates. In vitro regulation of [3H]imipramine binding by chloride was intact. Determination of the concentrations of 5-hydroxytryptamine (serotonin) and 5-hydroxyindoleacetic acid in hypothalamus and frontal cortex indicated no apparent age-related changes in indole metabolism. The age-related increase in brain [3H]imipramine binding and impairment in the in vitro regulation of binding by ions are similar to changes observed previously in aged mouse brain. The increase in brain antidepressant binding sites is discussed in relationship to other indices of brain serotonergic function in aging and to the relationship of [3H]imipramine binding and depression.     

Apparent resilience of a declining Afro‐Palaearctic migrant to forest loss on the wintering grounds

Populations of long‐distance migrant birds are declining but it is unknown what role land cover change in non‐breeding areas may be playing in this process. Using compositional analysis, we assessed habitat selection by one such migrant, the Wood Warbler Phylloscopus sibilatrix, at a wintering site in the forest–savannah transition zone in Eastern Region, Ghana. There was a preference for forest, a habitat that is in marked decline at this site. Annual habitat mapping revealed that the area of forest declined by 26% between 2011/12 and 2013/14, mainly through clearance for conversion to arable land. Numbers of birds changed throughout the season, but despite the reduction in the preferred forest habitat, there was no change in the total number of birds recorded at the site over the study period. The number of birds recorded at a point was positively related to the proportion of cleared land, plantation and, to a lesser extent, dense forest within 100 m. Investigation of the fine‐scale habitat preferences of radiotagged Wood Warblers suggested that there was an optimum number of trees, around 66–143 per hectare, at which estimated probability of occupancy was 0.5, falling to a probability of 0.2 at 25 trees per hectare. We suggest that Wood Warblers may be buffered against the loss of forest habitat by their ability to utilize degraded habitats, such as well‐wooded farmland, that still retain a substantial number of trees. However, the continued loss of trees, from both forest and farmland is ultimately likely to have a negative impact on wintering Wood Warblers in the long‐term.     

Overview,opportunities and outlook for Australian spiny lobster fisheries

Australia’s lobster fisheries are relatively small in volume (9500t) compared with global production (289,000t), but are the country’s most valuable in terms of both overall production and value of export (2014 Gross Value of Production of $610 million AUD). Further, they support commercial, recreational and indigenous fishers along most of the continent’s coastline. Here we review similarities and key differences between these lobster fisheries, based on biological characteristics, fishery data collection, assessment and management methods, and supply chain considerations. A diverse range of palinurid lobsters occur in Australia, but only three genera, distributed across eight different management jurisdictions, support significant fisheries. Catches of western rock lobster Panulirus cygnus dominate landings (61%), followed by southern rock lobster Jasus edwardsii, tropical lobster Panulirus ornatus and the eastern rock lobster Sagmariasus verreauxi. Large-scale environmental influences such as climate change are impacting on these fisheries in similar or different ways forcing new management and raising the need for greater resilience in current supply chains. Although these are separate fisheries, the integrated nature of the dominant Chinese export markets suggests potentially important economic and market-related interactions. Our overview highlights the critical role of continued monitoring of recruitment pulses, in combination with robust harvest strategies, to ensure that harvests respond adequately and fisheries achieve biological and economic sustainability. Approaches that also include socio-cultural considerations (triple bottom line) are important given many fisheries include indigenous Australians. Our integrated analysis of Australian lobster fisheries highlights differences and similarities with spiny lobster fisheries worldwide and lessons from opportunities, including adapting to new free trade agreements, enhancing the reputation of wild lobsters as a whole, sharing expertise, and better alignment of supply and demand.     

An Integrated Experimental and Computational Approach for Characterizing the Kinetics and Mechanism of Triadimefon Racemization

Enantiomers of chiral molecules commonly exhibit differing pharmacokinetics and toxicities, which can introduce significant uncertainty when evaluating biological and environmental fates and potential risks to humans and the environment. However, racemization (the irreversible transformation of one enantiomer into the racemic mixture) and enantiomerization (the reversible conversion of one enantiomer into the other) are poorly understood. To better understand these processes, we investigated the chiral fungicide, triadimefon, which undergoes racemization in soils, water, and organic solvents. Nuclear magnetic resonance (NMR) and gas chromatography / mass spectrometry (GC/MS) techniques were used to measure the rates of enantiomerization and racemization, deuterium isotope effects, and activation energies for triadimefon in H₂O and D₂O. From these results we were able to determine that: 1) the alpha‐carbonyl carbon of triadimefon is the reaction site; 2) cleavage of the C‐H (C‐D) bond is the rate‐determining step; 3) the reaction is base‐catalyzed; and 4) the reaction likely involves a symmetrical intermediate. The B3LYP/6–311 + G** level of theory was used to compute optimized geometries, harmonic vibrational frequencies, nature population analysis, and intrinsic reaction coordinates for triadimefon in water and three racemization pathways were hypothesized. This work provides an initial step in developing predictive, structure‐based models that are needed to identify compounds of concern that may undergo racemization. Chirality 28:633–641, 2016. © 2016 Wiley Periodicals, Inc.