Protein kinase A RII-like (R2D2) proteins exhibit differential localization and AKAP interaction

A-kinase anchoring proteins (AKAPs) bind to protein kinase A (PKA) via an amphipathic helix domain that interacts with a dimerization/docking domain on the regulatory (R) subunit of PKA. Four other mammalian proteins (ROPN1, ASP, SP17, and CABYR) also contain a highly conserved RII dimerization/docking (R2D2) domain, suggesting all four proteins may interact with all AKAPs in a manner similar to RII. All four of these proteins were originally detected in the flagellum of mammalian sperm. In this report, we demonstrate that all four R2D2 proteins are expressed in a wide variety of tissues and three of the proteins SP17, CABYR, and ASP are located in motile cilia of human bronchus and fallopian tubes. In addition, we detect SP17 in primary cilia. We also provide evidence that ROPN1 and ASP bind to a variety of AKAPs and this interaction can be disrupted with anchoring inhibitor peptides. The interaction of SP17 and CABYR with AKAPs appears to be much more limited. None of the R2D2 proteins appears to bind cAMP, a fundamental characteristic of the regulatory subunits of PKA. These observations suggest that R2D2 proteins utilize docking interactions with AKAPs to accomplish their function of regulating cilia and flagella. Based on location, affinity for AKAPs and lack of affinity for cAMP, it appears that each R2D2 protein has a unique role in this process.     

Superresolution imaging reveals structural features of EB1 in microtubule plus-end tracking

Visualization of specific molecules and their interactions in real time and space is essential to delineate how cellular dynamics and the signaling circuit are orchestrated. Spatial regulation of conformational dynamics and structural plasticity of protein interactions is required to rewire signaling circuitry in response to extracellular cues. We introduce a method for optically imaging intracellular protein interactions at nanometer spatial resolution in live cells, using photoactivatable complementary fluorescent (PACF) proteins. Subsets of complementary fluorescent protein molecules were activated, localized, and then bleached; this was followed by the assembly of superresolution images from aggregate position of sum interactive molecules. Using PACF, we obtained precise localization of dynamic microtubule plus-end hub protein EB1 dimers and their distinct distributions at the leading edges and in the cell bodies of migrating cells. We further delineated the structure–function relationship of EB1 by generating EB1-PACF dimers (EB1^wt:EB1^wt, EB1^wt:EB1^mt, and EB1^mt:EB1^mt) and imaging their precise localizations in culture cells. Surprisingly, our analyses revealed critical role of a previously uncharacterized EB1 linker region in tracking microtubule plus ends in live cells. Thus PACF provides a unique approach to delineating spatial dynamics of homo- or heterodimerized proteins at the nanometer scale and establishes a platform to report the precise regulation of protein interactions in space and time in live cells.     

The fall and rise of the Oblong woodsia in Britain

Summary

The vegetation of a small spoil heap at Jerah mine near Stirling was classified as species-rich acid grassland, an open barren-soil community with a high frequency of the moss Pohlia nutans, and a group which showed intermediate characteristics. Copper was present in the soil in potentially toxic concentrations and when the vegetation data were ordinated the axes showed highly significant correlations with soil copper and loss-on-ignition. The results suggest that copper is a major determinant of the spoil-heap vegetation, and contrast sharply with those reported for the Burn of Sorrow mine, a site with many superficial similarities.     

Overcoming the constraints of long range radio telemetry from animals: getting more useful data from smaller packages

Many species carry out their most interesting activities wherethey cannot readily be observed or monitored. Marine mammalsare extreme among this group, accomplishing their most astoundingactivities both distant from land and deep in the sea. Collection,storage and transmission of data about these activities areconstrained by the energy requirements and size of the recordingloggers and transmitters. The more bits of information collected,stored and transmitted, the more battery is required and thelarger the tag must be. We therefore need to be selective aboutthe information we collect, while maintaining detail and fidelity.To accomplish this in the study of marine mammals, we have designed"intelligent" data logger/transmitters that provide context-drivendata compression, data relay, and automated data base storage.We later combine these data with remotely sensed environmentalinformation and other oceanographic data sets to recreate theenvironmental context for the animal's activity, and we displaythe combined data using computer animation techniques. In thisway, the system can provide near real time "observation" ofanimal behavior and physiology from the remotest parts of theglobe.     

Lack of evidence for inhibitory processes in over-selectivity

Stimulus over-selectivity can be defined as control over behavior being exerted by one aspect of the environment at the expense of other equally salient aspects of the environment, and is a common problem for discrimination learning under conditions of cognitive strain, and in intellectual disorders. Non-clinical participants exposed to a concurrent task load were trained and tested on a two-component trial-and-error discrimination task to investigate whether inhibition plays a role in producing under-selectivity by using both summation and retardation tests. Experiment 1 found evidence for the over-selectivity effect, and replicated the finding that revaluation of a previously over-selected stimulus allows emergence of control by a previous under-selected stimulus, despite the latter stimulus receiving no direct conditioning. The under-selected cue was not found to gain any conditioned inhibitory status, as reflected by summation (Experiment 2), and retardation (Experiment 3), tests. The results extend the literature explaining over-selectivity as a post-acquisition failure.