Identification of a new genomic hot spot of evolutionary diversification of protein function.

Establishment of phylogenetic relationships remains a challenging task because it is based on computational analysis of genomic hot spots that display species-specific sequence variations. Here, we identify a species-specific thymine-to-guanine sequence variation in the Glrb gene which gives rise to species-specific splice donor sites in the Glrb genes of mouse and bushbaby. The resulting splice insert in the receptor for the inhibitory neurotransmitter glycine (GlyR) conveys synaptic receptor clustering and specific association with a particular synaptic plasticity-related splice variant of the postsynaptic scaffold protein gephyrin. This study identifies a new genomic hot spot which contributes to phylogenetic diversification of protein function and advances our understanding of phylogenetic relationships.

Attributing trends in extremely hot days to changes in atmospheric dynamics

This paper presents a method for attributing regional trends in the frequency of extremely hot days (EHDs) to changes in the frequency of the atmospheric patterns that characterize such extraordinary events. The study is applied to mainland Spain and the Balearic Islands for the extended summers of the period 1958–2008, where significant and positive trends in maximum temperature (Tx) have been reported during the second half of the past century. First, the study area was split into eight regions attending to their different temporal variability of the daily Tx series obtained from the Spain02 gridded data set using a clustering procedure. Second, the large-scale atmospheric situations causing EHDs are defined by circulation types (CTs). The obtainment of the CTs differs from the majority of CT classifications proposed in the literature. It is based on regional series and on a previous characterization of the main atmospheric situations obtained using only some days classified as extremes in the different regions. Three different atmospheric fields (SLP, T850, and Z500) from ECMWF reanalysis and analysis data and combinations of them (SLP–T850, SLP–Z500, and T850–Z500) are used to produce six different CT classifications. Subsequently, links between EHD occurrence in the different regions and CT for all days have been established. Finally, a simple model to relate the trends in EHDsfor each region to the changes in the CT frequency appearance has been formulated. Most regions present positive and significant trends in the occurrence of EHDs. The CT classifications using two variables perform better. In particular, SLP–T850 is the best for characterizing the atmospheric situations leading to EHD occurrences for most of the regions. Only a small number of CTs have significant trends in their frequency and are associated with high efficiency causing EHD occurrences in most regions simultaneously, especially in the northern and central regions. Attribution results show that changes in circulation can only explain some part of the regional EHD trends. The percentage of the trend attributable to changes in atmospheric dynamics varies from 15 to 50 %, depends on the region and is sensitive to the selected large-scale variables.

Electron cryomicroscopy of E. coli reveals filament bundles involved in plasmid DNA segregation.

Bipolar elongation of filaments of the bacterial actin homolog ParM drives movement of newly replicated plasmid DNA to opposite poles of a bacterial cell. We used a combination of vitreous sectioning and electron cryotomography to study this DNA partitioning system directly in native, frozen cells. The diffraction patterns from overexpressed ParM bundles in electron cryotomographic reconstructions were used to unambiguously identify ParM filaments in Escherichia coli cells. Using a low-copy number plasmid encoding components required for partitioning, we observed small bundles of three to five intracellular ParM filaments that were situated close to the edge of the nucleoid. We propose that this may indicate the capture of plasmid DNA within the periphery of this loosely defined, chromosome-containing region.

Development of a Novel Green Fluorescent Protein-Based Binding Assay to Study the Association of Plakins with Intermediate Filament Proteins.

Protein-protein interactions are fundamental for most biological processes, such as the formation of cellular structures and enzymatic complexes or in signaling pathways. The identification and characterization of protein-protein interactions are therefore essential for understanding the mechanisms and regulation of biological systems. The organization and dynamics of the cytoskeleton, as well as its anchorage to specific sites in the plasma membrane and organelles, are regulated by the plakins. These structurally related proteins anchor different cytoskeletal networks to each other and/or to other cellular structures. The association of several plakins with intermediate filaments (IFs) is critical for maintenance of the cytoarchitecture. Pathogenic mutations in the genes encoding different plakins can lead to dramatic manifestations, occurring principally in the skin, striated muscle, and/or nervous system, due to cytoskeletal disorganization resulting in abnormal cell fragility. Nevertheless, it is still unclear how plakins bind to IFs, although some general rules are slowly emerging. We here describe in detail a recently developed protein-protein fluorescence binding assay, based on the production of recombinant proteins tagged with green fluorescent protein (GFP) and their use as fluid-phase fluorescent ligands on immobilized IF proteins. Using this method, we have been able to assess the ability of C-terminal regions of GFP-tagged plakin proteins to bind to distinct IF proteins and IF domains. This simple and sensitive technique, which is expected to facilitate further studies in this area, can also be potentially employed for any kind of protein-protein interaction studies.