Project-based work and sustainable development : a comparative case study of cultural animation projects

Currently, a growing interest in the issues related to sustainable development can be observed, with the role of culture in stimulating this development increasing simultaneously. Nevertheless, the function and meanings of culture for sustainable development, as well as culture in the context of sustainable development, have so far remained under-emphasised and under-theorised. For this reason, in this paper we will look at practical examples of culture and sustainable development combined. The undertaken research problem explores the project-based work in the field of cultural animation, and its impact on the pursuit of the objectives of sustainable development at the local level. Two case studies of Polish organisations involved in cultural animation activities have been analysed herein. Based on the research results, we showed that cultural animation is an important tool for enabling local communities to achieve sustainable development. What is also important is the fact that cultural animation activities often take the form of project-based work, which significantly affects the methodology and extent of their implementation. Therefore, in this article we also point to the relationship between the management of animation projects and the idea of sustainable development, emphasising both advantages and disadvantages thereof

Phosphorylation of spore coat proteins by a family of atypical protein kinases

The modification of proteins by phosphorylation occurs in all life forms and is catalyzed by a large superfamily of enzymes known as protein kinases. We recently discovered a family of secretory pathway kinases that phosphorylate extracellular proteins. One member, family with sequence similarity 20C (Fam20C), is the physiological Golgi casein kinase. While examining distantly related protein sequences, we observed low levels of identity between the spore coat protein H (CotH), and the Fam20C-related secretory pathway kinases. CotH is a component of the spore in many bacterial and eukaryotic species, and is required for efficient germination of spores in Bacillus subtilis; however, the mechanism by which CotH affects germination is unclear. Here, we show that CotH is a protein kinase. The crystal structure of CotH reveals an atypical protein kinase-like fold with a unique mode of ATP binding. Examination of the genes neighboring cotH in B. subtilis led us to identify two spore coat proteins, CotB and CotG, as CotH substrates. Furthermore, we show that CotH-dependent phosphorylation of CotB and CotG is required for the efficient germination of B. subtilis spores. Collectively, our results define a family of atypical protein kinases and reveal an unexpected role for protein phosphorylation in spore biology

A Novel Protein Kinase-Like Domain in a Selenoprotein, Widespread in the Tree of Life

Selenoproteins serve important functions in many organisms, usually providing essential oxidoreductase enzymatic activity, often for defense against toxic xenobiotic substances. Most eukaryotic genomes possess a small number of these proteins, usually not more than 20. Selenoproteins belong to various structural classes, often related to oxidoreductase function, yet a few of them are completely uncharacterised

CLCAs - a family of metalloproteases of intriguing phylogenetic distribution and with cases of substituted catalytic sites.

The zinc-dependent metalloproteases with His-Glu-x-x-His (HExxH) active site motif, zincins, are a broad group of proteins involved in many metabolic and regulatory functions, and found in all forms of life. Human genome contains more than 100 genes encoding proteins with known zincin-like domains. A survey of all proteins containing the HExxH motif shows that approximately 52% of HExxH occurrences fall within known protein structural domains (as defined in the Pfam database). Domain families with majority of members possessing a conserved HExxH motif include, not surprisingly, many known and putative metalloproteases. Furthermore, several HExxH-containing protein domains thus identified can be confidently predicted to be putative peptidases of zincin fold. Thus, we predict zincin-like fold for eight uncharacterised Pfam families. Besides the domains with the HExxH motif strictly conserved, and those with sporadic occurrences, intermediate families are identified that contain some members with a conserved HExxH motif, but also many homologues with substitutions at the conserved positions. Such substitutions can be evolutionarily conserved and non-random, yet functional roles of these inactive zincins are not known. The CLCAs are a novel zincin-like protease family with many cases of substituted active sites. We show that this allegedly metazoan family has a number of bacterial and archaeal members. An extremely patchy phylogenetic distribution of CLCAs in prokaryotes and their conserved protein domain composition strongly suggests an evolutionary scenario of horizontal gene transfer (HGT) from multicellular eukaryotes to bacteria, providing an example of eukaryote-derived xenologues in bacterial genomes. Additionally, in a protein family identified here as closely homologous to CLCA, the CLCA_X (CLCA-like) family, a number of proteins is found in phages and plasmids, supporting the HGT scenario

A Combined Methods of Senile Trees Inventory in Sustainable Urban Greenery Management on the Example of the City of Sandomierz (Poland)

The sustainable management of urban greenery consists, among others, of the inventory, valuation, and protection of trees of monumental size. This article presents the results of the inspection of 13 large trees growing in the city of Sandomierz, located in south-eastern Poland. The examined specimens belong to five species: Norway maple (Acer platanoides L.), common ash (Fraxinus excelsior L.), white poplar (Populus alba L.), English oak (Quercus robur L.), and small-leaved lime (Tilia cordata Mill.). The health condition of the trees was assessed using acoustic and electrical tomography, as well as chlorophyll fluorescence tests. Diagnostics employing sound waves and electrical resistivity were crucial in assessing tree health. The data based on chlorophyll fluorescence confirmed the results obtained during tomographic examinations. It was an innovative combination of three non-invasive methods of examining the health condition of trees and their valuation. Economic valuation allows us to reduce to common denominator issues that are often difficult to decide due to different perspectives—expressing the economic value of trees. Calculating the value of trees allowed us to show the city’s inhabitants the value of trees that are of monumental size. Thanks to the cooperation of scientists with the city authorities, an economic plan for trees of monumental size was created, distinguished by an individualized and holistic approach to each specimen covered by the study. The database prepared has a chance to become an effective management instrument used by environmental protection authorities and a source of knowledge and education for the city’s inhabitants

Multiple sequence alignments of selected FAM69 proteins.

<p><i>Left:</i> alignment of the kinase domain, covering the region 83–422 of FAM69A. The sequences are aligned using Promals3D (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066427#s4" target="_blank">Methods</a>). Secondary structure prediction for human shown for selected proteins, for the solved structures actual secondary structure shown. Secondary structure elements named as in PKA <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066427#pone.0066427-Knighton1" target="_blank">[87]</a>. Locations of predicted key catalytic residues shown, in standard PKA numbering (e.g. D166), as well as the ATP-binding loop (GxGxxG). SwissProt identifiers shown for human sequences, otherwise, NCBI GI identifiers shown together with abbreviations of species names: Sp: sea urchin <i>Strongylocentrotus purpuratus, Nv:</i> sea cucomber <i>Nematostella vectens, Dm:</i> fruit fly <i>Drosophila melanogaster; Mm: Mus musculus; Ce: Caenorhabditis elegans, Co: Capsaspora owczarzaki; S sp: Salpingoeca sp. ATCC 50818</i>. Also shown selected close kinase homologues (PKDCC and SGK196) as well as sequences of selected kinases of known structures. Numbers in brackets indicate numbers of residues omitted from the alignment (shown only for the 1 cdk, 3 sxs structures, and for human FAM69A, DIA1, PKDCC and SGK196 sequences). R and C characters on black background above the alignment indicate the regulatory and catalytic spine residues, respectively <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066427#pone.0066427-Taylor2" target="_blank">[22]</a>. The location of the the EF-hand motif shown, the motif itself is excised from the alignment and shown on the right. <i>Right:</i> alignment of the EF-hand region (corresponding to the region 165–199 of human FAM69A). Also shown EF-hand regions of human calmodulin and the 2PMY region used for model building.</p

Kinase motifs.

<p>Sequence logos for selected kinase motifs in the FAM69 family. <i>Top:</i> the β-4 - β-5 region. <i>Middle row, left:</i> the predicted active site region (corresponding to D166 and N171 in PKA). <i>Middle row, right</i>- the Mg²⁺ - binding motif (corresponding to the DFG motif in PKA). <i>Bottom, left:</i> predicted helix α-F. <i>Bottom, right:</i> predicted helix α-G.</p

A novel predicted ADP-ribosyltransferase-like family conserved in eukaryotic evolution

The presence of many completely uncharacterized proteins, even in well-studied organisms such as humans, seriously hampers full understanding of the functioning of the living cells. ADP-ribosylation is a common post-translational modification of proteins; also nucleic acids and small molecules can be modified by the covalent attachment of ADP-ribose. This modification, important in cellular signalling and infection processes, is usually executed by enzymes from the large superfamily of ADP-ribosyltransferases (ARTs). Here, using bioinformatics approaches, we identify a novel putative ADP-ribosyltransferase family, conserved in eukaryotic evolution, with a divergent active site. The hallmark of these proteins is the ART domain nestled between flanking leucine-rich repeat (LRR) domains. LRRs are typically involved in innate immune surveillance. The novel family appears as putative novel ADP-ribosylationrelated actors, most likely pseudoenzymes. Sequence divergence and lack of clearly detectable ''classical'' ART active site suggests the novel domains are pseudoARTs, yet atypical ART activity, or alternative enzymatic activity cannot be excluded. We propose that this family, including its human member LRRC9, may be involved in an ancient defense mechanism, with analogies to the innate immune system, and coupling pathogen detection to ADP-ribosyltransfer or other signalling mechanisms

Structure model of the kinase domain of human FAM69A.

<p><i>Top left:</i> model coloured by MetaMQAP model quality score (blue: good quality, red: poor quality). On left, the EF-hand motif is shown in yellow. <i>Top right:</i> as in Fig. 5 (top left), model coloured by sequence: from dark blue (N-terminus) to dark red (C-terminus). <i>Bottom:</i> close-up of the predicted active site with ATP molecule bound. Side chains of key predicted active site residues shown: D294 (PKA numbering: 166), N299 (171), D312 (184), also the two cysteines near the predicted active site that may form a S-S bridge: C293 and C331.</p