A Ubl/ubiquitin switch in the activation of Parkin

Mutations in Parkin and PINK1 cause an inherited early‐onset form of Parkinson's disease. The two proteins function together in a mitochondrial quality control pathway whereby PINK1 accumulates on damaged mitochondria and activates Parkin to induce mitophagy. How PINK1 kinase activity releases the auto‐inhibited ubiquitin ligase activity of Parkin remains unclear. Here, we identify a binding switch between phospho‐ubiquitin (pUb) and the ubiquitin‐like domain (Ubl) of Parkin as a key element. By mutagenesis and SAXS, we show that pUb binds to RING1 of Parkin at a site formed by His302 and Arg305. pUb binding promotes disengagement of the Ubl from RING1 and subsequent Parkin phosphorylation. A crystal structure of Parkin Δ86–130 at 2.54 Å resolution allowed the design of mutations that specifically release the Ubl domain from RING1. These mutations mimic pUb binding and promote Parkin phosphorylation. Measurements of the E2 ubiquitin‐conjugating enzyme UbcH7 binding to Parkin and Parkin E3 ligase activity suggest that Parkin phosphorylation regulates E3 ligase activity downstream of pUb binding.     

Mitigation of salinity-induced negative impact on the growth and yield of wheat by plant growth-promoting rhizobacteria in naturally saline conditions

Salinity is one of the major environmental threats for successful crop production, hampering plant growth due to the osmotic effect and nutritional and hormonal imbalances. The application of naturally occurring plant growth-promoting rhizobacteria (PGPR) is an emerging technology aimed at ameliorating the negative impact of salinity. However, the results obtained in the laboratory can sometimes not be reproduced in the field. The aim of the study reported here was to evaluate the effect of PGPR inoculation on seed germination in a saline environment under axenic conditions and on enhancement of the growth and yield of wheat under natural salt-affected field conditions. Wheat seeds were inoculated with pre-isolated strains of Pseudomonas putida, Enterobacter cloacae, Serratia ficaria, and Pseudomonas fluorescens and sown at different salinity levels (1, 2, 3, 6, 9, 12, 15 dS m^-1). Inoculation with these strains was found to enhance the germination percentage, germination rate, and index of wheat seeds up to 43, 51, and 123 %, respectively, over the uninoculated control at the highest salinity level. The potential of these PGPR for improving the growth and yield of wheat was also evaluated at two natural salt-affected sites. Inoculation with PGPR resulted a significant increase in the growth and yield parameters of wheat at both sites. The inoculated plants also improved the nutrient status of the wheat plants. The inoculated plants had low sodium and high nitrogen, phosphorus, and potassium contents. Our results show that such rhizobacterial strains may be used as an effective tool for enhancing plant growth under salinity stress and for maximizing the utilization of salt-affected soils.     

iDRP-PseAAC: Identification of DNA Replication Proteins Using General PseAAC and Position Dependent Features

International Journal of Peptide Research and Therapeutics - DNA replication is one of the specific processes to be considered in all the living organisms, specifically eukaryotes. The prevalence...     

Construct social‐behavioral association network to study management impact on waterbirds community ecology using digital video recording cameras

Studying social‐behavior and species associations in ecological communities is challenging because it is difficult to observe the interactions in the field. Animal behavior is especially difficult to observe when selection of habitat and activities are linked to energy costs of long‐distance movement. Migrating communities tend to be resource specific and prefer environments that offer more suitability for coexisting in a shared space and time. Given the recent advances in digital technologies, digital video recording systems are gaining popularity in wildlife research and management. We used digital video recording cameras to study social interactions and species–habitat linkages for wintering waterbirds communities in shared habitats. Examining over 8,640 hr of video footages, we built tetrapartite social‐behavioral association network of wintering waterbirds over habitat (n = 5) selection events in sites with distinct management regimes. We analyzed these networks to identify hub species and species role in activity persistence, and to explore the effects of hydrological regime on these network characteristics. Although the differences in network attributes were not significant at treatment level (p = .297) in terms of network composition and keystone species composition, our results indicated that network attributes were significantly different (p = .000, r ² = .278) at habitat level. There were evidences suggesting that the habitat quality was better at the managed sites, where the formed networks had more species, more network nodes and edges, higher edge density, and stronger intra‐ and inter‐species interactions. In addition, we also calculated the species interaction preference scores (SIPS) and behavioral interaction preference scores (BIPS) of each network. The results showed that species synchronize activities in shared space for temporal niche partitioning in order to avoid or minimize any potential competition for shared space. Our social network analysis (SNA) approach is likely to provide a practical use for ecosystem management and biodiversity conservation.     

Proviral Integration Site for Moloney Murine Leukemia Virus (PIM) Kinases Promote Human T Helper 1 Cell Differentiation

The differentiation of human primary T helper 1 (Th1) cells from naïve precursor cells is regulated by a complex, interrelated signaling network. The identification of factors regulating the early steps of Th1 cell polarization can provide important insight in the development of therapeutics for many inflammatory and autoimmune diseases. The serine/threonine-specific proviral integration site for Moloney murine leukemia virus (PIM) kinases PIM1 and PIM2 have been implicated in the cytokine-dependent proliferation and survival of lymphocytes. We have established that the third member of this family, PIM3, is also expressed in human primary Th cells and identified a new function for the entire PIM kinase family in T lymphocytes. Although PIM kinases are expressed more in Th1 than Th2 cells, we demonstrate here that these kinases positively influence Th1 cell differentiation. Our RNA interference results from human primary Th cells also suggest that PIM kinases promote the production of IFNγ, the hallmark cytokine produced by Th1 cells. Consistent with this, they also seem to be important for the up-regulation of the critical Th1-driving factor, T box expressed in T cells (T-BET), and the IL-12/STAT4 signaling pathway during the early Th1 differentiation process. In summary, we have identified PIM kinases as new regulators of human primary Th1 cell differentiation, thus providing new insights into the mechanisms controlling the selective development of human Th cell subsets.     

Response of microbial communities to elevated thallium contamination in river sediments

Abstract

The study of microbial communities in river sediments contaminated by thallium (Tl) is necessary to achieve the information for in-situ microbially mediated bioremediation. However, little is known about the microbial community in Tl-contaminated river sediments. In the present study, we characterized the microbial community and their responses to Tl pollution in river sediments from the Tl-mineralized Lanmuchang area, Southwest Guizhou, China. Illumina sequencing of 16S rRNA amplicons revealed that over 40 phyla belong to the domain bacteria. In all samples, Proteobacteria, Cyanobacteria, and Actinobacteria were the most dominant phyla. Based on the UPGMA (Unweighted Pair Group Method with Arithmetic Mean) tree and PCoA (Principal Coordinates Analysis) analysis, microbial composition of each segment was distinct, indicating in-situ geochemical parameters (including Tl, sulfate, TOC, Eh, and pH) had influenced on the microbial communities. Moreover, canonical correspondence analysis (CCA) was employed to further elucidate the impact of geochemical parameters on the distribution of microbial communities in local river sediments. The results indicated that a number of microbial communities including Cyanobacteria, Spirochaete, Hydrogenophaga, and Acinetobacter were positively correlated with total Tl, suggesting potential roles of these microbes to Tl tolerance or to biogeochemical cycling of Tl. Our results suggested a reliable location for the microbial community’s diversity in the presence of high concentrations of Tl and might have a potential association for in-situ bioremediation strategies of Tl-contaminated river. Overall, in situ microbial community could provide a useful tool for monitoring and assessing geo-environmental stressors in Tl-polluted river sediments.     

Structure and kinetic characterization of human sperm-specific glyceraldehyde-3-phosphate dehydrogenase, GAPDS

hGAPDS (human sperm-specific glyceraldehyde-3-phosphate dehydrogenase) is a glycolytic enzyme essential for the survival of spermatozoa, and constitutes a potential target for non-hormonal contraception. However, enzyme characterization of GAPDS has been hampered by the difficulty in producing soluble recombinant protein. In the present study, we have overexpressed in Escherichia coli a highly soluble form of hGAPDS truncated at the N-terminus (hGAPDSΔN), and crystallized the homotetrameric enzyme in two ligand complexes. The hGAPDSΔN-NAD+-phosphate structure maps the two anion-recognition sites within the catalytic pocket that correspond to the conserved Ps site and the newly recognized Pi site identified in other organisms. The hGAPDSΔN-NAD+-glycerol structure shows serendipitous binding of glycerol at the Ps and new Pi sites, demonstrating the propensity of these anion-recognition sites to bind non-physiologically relevant ligands. A comparison of kinetic profiles between hGAPDSΔN and its somatic equivalent reveals a 3-fold increase in catalytic efficiency for hGAPDSΔN. This may be attributable to subtle amino acid substitutions peripheral to the active centre that influence the charge properties and protonation states of catalytic residues. Our data therefore elucidate structural and kinetic features of hGAPDS that might provide insightful information towards inhibitor development.     

Structural and biochemical characterization of the salicylyl-acyltranferase SsfX3 from a tetracycline biosynthetic pathway

SsfX3 is a GDSL family acyltransferase that transfers salicylate to the C-4 hydroxyl of a tetracycline intermediate in the penultimate step during biosynthesis of the anticancer natural product SF2575. The C-4 salicylate takes the place of the more common C-4 dimethylamine functionality, making SsfX3 the first acyltransferase identified to act on a tetracycline substrate. The crystal structure of SsfX3 was determined at 2.5 Å, revealing two distinct domains as follows: an N-terminal β-sandwich domain that resembles a carbohydrate-binding module, and a C-terminal catalytic domain that contains the atypical α/β-hydrolase fold found in the GDSL hydrolase family of enzymes. The active site lies at one end of a large open binding pocket, which is spatially defined by structural elements from both the N- and C-terminal domains. Mutational analysis in the putative substrate binding pocket identified residues from both domains that are important for binding the acyl donor and acceptor. Furthermore, removal of the N-terminal carbohydrate-binding module-like domain rendered the stand-alone α/β-hydrolase domain inactive. The additional noncatalytic module is therefore proposed to be required to define the binding pocket and provide sufficient interactions with the spatially extended tetracyclic substrate. SsfX3 was also demonstrated to accept a variety of non-native acyl groups. This relaxed substrate specificity toward the acyl donor allowed the chemoenzymatic biosynthesis of C-4-modified analogs of the immediate precursor to the bioactive SF2575; these were used to assay the structure activity relationships at the C-4 position.