Role of differential expression of streptococcal arginine deiminase in inhibition of fimA expression in Porphyromonas gingivalis

Streptococcus cristatus ArcA inhibits production of a major adhesin, FimA, in Porphyromonas gingivalis, a primary periodontal pathogen. In this study, we demonstrate the differential expression of arcA in two streptococcal species. The expression level of arcA in streptococci appears to be controlled by both cis and trans elements.     

Role of the Clp system in stress tolerance, biofilm formation, and intracellular invasion in Porphyromonas gingivalis

Clp proteases and chaperones are ubiquitous among prokaryotes and eukaryotes, and in many pathogenic bacteria the Clp stress response system is also involved in regulation of virulence properties. In this study, the roles of ClpB, ClpC, and ClpXP in stress resistance, homotypic and heterotypic biofilm formation, and intracellular invasion in the oral opportunistic pathogen Porphyromonas gingivalis were investigated. Absence of ClpC and ClpXP, but not ClpB, resulted in diminished tolerance to high temperatures. Response to oxidative stress was not affected by the loss of any of the Clp proteins. The clpC and clpXP mutants demonstrated elevated monospecies biofilm formation, and the absence of ClpXP also enhanced heterotypic P. gingivalis-Streptococcus gordonii biofilm formation. All clp mutants adhered to gingival epithelial cells to the same level as the wild type; however, ClpC and ClpXP were found to be necessary for entry into host epithelial cells. ClpB did not play a role in entry but was required for intracellular replication and survival. ClpXP negatively regulated the surface exposure of the minor fimbrial (Mfa) protein subunit of P. gingivalis, which stimulates biofilm formation but interferes with epithelial cell entry. Collectively, these results show that the Clp protease complex and chaperones control several processes that are important for the colonization and survival of P. gingivalis in the oral cavity.     

Structure–function relationships in the bifunctional ferrisiderophore FpvA receptor from Pseudomonas aeruginosa

FpvA is the primary outer membrane transporter required for iron acquisition via the siderophore pyoverdine (Pvd) in Pseudomonas aeruginosa. FpvA, like other ferrisiderophore transporters, consists of a membrane-spanning β-barrel occluded by a plug domain. The β-strands of the barrel are connected by large extracellular loops and periplasmic turns. Like some other TonB-dependent transporters, FpvA has a periplasmic domain involved in a signalling cascade that regulates expression of genes required for ferrisiderophore transport. Here, the structures of FpvA in different loading states are analysed in light of mutagenesis data. This analysis highlights the roles of different protein domains in Pvd-Fe uptake and the signalling cascade and reveals a strong correlation between Pvd-Fe transport and activation of the signalling cascade. It is likely that conclusions drawn for FpvA will be relevant to other TonB-dependent ferrisiderophore transport and signalling proteins.     

Proteomic profiling of neuronal mitochondria reveals modulators of synaptic architecture

Background

Neurons are highly polarized cells consisting of three distinct functional domains: the cell body (and associated dendrites), the axon and the synapse. Previously, it was believed that the clinical phenotypes of neurodegenerative diseases were caused by the loss of entire neurons, however it has recently become apparent that these neuronal sub-compartments can degenerate independently, with synapses being particularly vulnerable to a broad range of stimuli. Whilst the properties governing the differential degenerative mechanisms remain unknown, mitochondria consistently appear in the literature, suggesting these somewhat promiscuous organelles may play a role in affecting synaptic stability. Synaptic and non-synaptic mitochondrial subpools are known to have different enzymatic properties (first demonstrated by Lai et al., 1977). However, the molecular basis underpinning these alterations, and their effects on morphology, has not been well documented.

Methods

The current study has employed electron microscopy, label-free proteomics and in silico analyses to characterize the morphological and biochemical properties of discrete sub-populations of mitochondria. The physiological relevance of these findings was confirmed in-vivo using a molecular genetic approach at the Drosophila neuromuscular junction.

Results

Here, we demonstrate that mitochondria at the synaptic terminal are indeed morphologically different to non-synaptic mitochondria, in both rodents and human patients. Furthermore, generation of proteomic profiles reveals distinct molecular fingerprints – highlighting that the properties of complex I may represent an important specialisation of synaptic mitochondria. Evidence also suggests that at least 30% of the mitochondrial enzymatic activity differences previously reported can be accounted for by protein abundance. Finally, we demonstrate that the molecular differences between discrete mitochondrial sub-populations are capable of selectively influencing synaptic morphology in-vivo. We offer several novel mitochondrial candidates that have the propensity to significantly alter the synaptic architecture in-vivo.

Conclusions

Our study demonstrates discrete proteomic profiles exist dependent upon mitochondrial subcellular localization and selective alteration of intrinsic mitochondrial proteins alters synaptic morphology in-vivo.

     

Regulation of olfactomedin 4 by Porphyromonas gingivalis in a community context

At mucosal barriers, the virulence of microbial communities reflects the outcome of both dysbiotic and eubiotic interactions with the host, with commensal species mitigating or potentiating the action of pathogens. We examined epithelial responses to the oral pathogen Porphyromonas gingivalis as a monoinfection and in association with a community partner, Streptococcus gordonii. RNA-Seq of oral epithelial cells showed that the Notch signaling pathway, including the downstream effector olfactomedin 4 (OLFM4), was differentially regulated by P. gingivalis alone; however, regulation was overridden by S. gordonii. OLFM4 was required for epithelial cell migratory, proliferative and inflammatory responses to P. gingivalis. Activation of Notch signaling was induced through increased expression of the Notch1 receptor and the Jagged1 (Jag1) agonist. In addition, Jag1 was released in response to P. gingivalis, leading to paracrine activation. Following Jag1-Notch1 engagement, the Notch1 extracellular domain was cleaved by P. gingivalis gingipain proteases. Antagonism by S. gordonii involved inhibition of gingipain activity by secreted hydrogen peroxide. The results establish a novel mechanism by which P. gingivalis modulates epithelial cell function which is dependent on community context. These interrelationships have relevance for innate inflammatory responses and epithelial cell fate decisions in oral health and disease.Subject terms: Microbial ecology, Microbial ecology     

Vegetation along hydrologic, edaphic, and geochemical gradients in a high-elevation poor fen in Canaan Valley, West Virginia

Wetland plant community composition and pattern are regulated by a host of abiotic/environmental gradients and biotic factors. We used multivariate analyses to classify wetland plant communities and determine the relation of hydrologic, edaphic, and geochemical gradients on community composition and spatial distribution among 18 vegetation, hydrology, and soil sampling points in Abe Run, a botanically unique poor fen in northeastern West Virginia. We also examined the interactions of disturbance with the physical environment and species composition. A total of 179 vascular plant species were identified from sample plots. Vegetative composition and structure, dominant soil texture, and hydrology lacked the distinct concentric zonation of northern peatlands; instead, all were patchy and varied considerably over short distances. Graminoid-forb meadows with primarily silt-loam mineral horizons, greater depth to groundwater, and fewer days of inundation characterized the lower reaches of the wetland. These plots were more acidic, with absent or shallow O horizons, and lower concentrations of soil base cations (Ca, Mg, K). In the upper reaches of the wetland, mixed herb-shrub-tree dominated communities were structurally and compositionally more complex; here, organic horizons were much more prominent, peat depth ranged from 80 to 100 cm, and the average depth to water table was 10 cm less than for wells in the lower reaches of the wetland. Plots from upper transects (3–6) tended to have more shrub and tree cover, and higher concentrations of soil base cations. Much of the variability among plots in the upper and lower reaches of the wetland are consistent with beaver inundation of a large portion of the wetland during the 1980s. Multiple-response permutation procedures verified the difference (P < 0.0001) between vegetation of plots in the lower and upper reaches of the watershed. Because fens are connected to groundwater, these habitats are particularly vulnerable to disturbances, particularly those that alter existing land use and land cover. Minimization of disturbances in the surrounding watershed, controls on an excessive deer population, removal of exotic, non-native species, and control on foot traffic are all integral to maintaining the integrity of this high-value wetland.     

Melanesian Kastom and Its Transformations

Editorial Board Note: In 2006, the Discipline Group of Anthropology and Sociology at The University of Western Australia hosted a symposium, ‘Anthropology in the West: 1956–2006’, to celebrate the 50th anniversary of the founding of the then Department of Anthropology by the late Ronald Berndt and his wife, Catherine, whose contributions to anthropology were a major focus of the symposium. Symposium participants also documented contributions made by UWA staff and students from Anthropology and Sociology within and beyond Australian social science. A notable theoretical focus on kastom in the work of Robert Tonkinson, who in 1984 succeeded Professor Berndt in the Departmental Chair, was the topic of Lamont Lindstrom's paper, published here in a fuller, revised and fully refereed version. We intend to publish further papers from the symposium in later issues of Anthropological Forum.

If Ronald and Catherine Berndt are ancestral spirits haunting Anthropology and Sociology at The University of Western Australia some 50 years after the founding of the Department of Anthropology, their student, Bob Tonkinson, still happily with us, is their intellectual descendant and institutional heir. As in the case of the Berndts, issues relating to social change, religion and values, and, in particular, the politics of tradition have loomed large in Tonkinson's career. I trace, in this retrospective essay, the rise to prominence of an anthropology of Melanesian tradition and, more specifically, Tonkinson's contribution, notably his analyses of tradition's ‘symbiotic’ relations with Christianity, its identity functions, its local versus its national significance, and its relations with evolving anthropological theories of culture in a shrinking world. Tonkinson's Vanuatu research, which began among Ambrym Island emigrants, has spanned, over the past forty years or so, many notable transformations: from New Hebrides to Vanuatu; from modernisation theory to world systems and globalisation; and from tradition to kastom.     

Survival and growth of native and exotic composites in response to a nutrient gradient

Mortality and performance (total biomass) of five exotic (weedy) and five native annual Asteraceae in Western Australia were compared along a mineral nutrient gradient. Mortality did not differ between exotic and native species but varied greatly within each group. The exotic species responded more positively to higher nutrient additions than the native species, indicating that the exotics might have a competitive advantage in a nutrient-enhanced situation (e.g., after fire). The concept of phenotypic inertia (PI), combining survival and performance over a range of environments, is introduced as a means of evaluating the range of growing conditions tolerated. The two species with highest PI (one exotic and one native) had the lowest mortality and uniformly moderate biomass for the nine nutrient treatments. The species with the lowest PI (a native) had complete failure at the two highest nutrient levels and low variable growth in the remainder. PI has potential as a measure of environmental tolerance although, in the present study, the exotic species were not clearly superior to the native species in their responses to a wide range of soil nutrient levels.     

Siderophore-mediated cell signalling in Pseudomonas aeruginosa: divergent pathways regulate virulence factor production and siderophore receptor synthesis

Under iron-limiting conditions, Pseudomonas aeruginosa produces a siderophore called pyoverdine. Pyoverdine is secreted into the extracellular environment where it chelates iron, and the resulting ferri-pyoverdine complexes are transported back into the bacteria by a cell surface receptor protein FpvA. Pyoverdine also acts as a signalling molecule inducing the production of three secreted virulence factors. Binding of ferri-pyoverdine to FpvA transduces a signal to the periplasmic part of the membrane-spanning antisigma factor FpvR. The signal is transmitted to the cytoplasmic part of FpvR, which controls the activity of an extracytoplasmic family (ECF) sigma factor protein PvdS. This results in the production of the virulence factors pyoverdine, exotoxin A and PrpL endoprotease. Here, we show that a second divergent branch of this signalling pathway regulates the production of the FpvA protein. FpvR negatively regulates the activity of a second ECF sigma factor, FpvI, which is required for the synthesis of FpvA, and the presence of ferri-pyoverdine greatly increases the activity of FpvI so that production of FpvA is induced. To the best of our knowledge, this is the first example of a branched signalling system of this sort and the first example of an antisigma factor protein (FpvR) that directly regulates the activities of two different ECF sigma factor proteins (PvdS and FpvI).     

Association of MHC class I and class II gene polymorphisms with vaccine or challenge response to Salmonella enteritidis in young chicks

Salmonella enteritidis (SE) is a worldwide source of salmonellosis in humans, caused mainly by consumption of contaminated poultry products. The major histocompatibility complex (MHC) class I and class II molecules, which function as antigen-presentation structures, are involved in both macrophage and dendritic cell immune response to Salmonella and may, therefore, play an important role in host resistance to infections. The chicken MHC class I and class II were investigated as candidate genes for immune response to SE. We characterized the complete MHC class I cDNA sequences for an outbred broiler line and four diverse highly inbred lines: two MHC-congenic Leghorn (G-B2 and G-B1), one Egyptian Fayoumi (M15.2), and one Spanish (Sp21.1), to define the allelic sequences within these lines, so that we might study the association between particular MHC polymorphisms and response to SE. The F1 offspring of outbred broiler sires crossed with three inbred lines (G-B1, G-B2, and Fayoumi) were evaluated as young chicks for either bacterial load in spleen and cecum after pathogenic SE inoculation or antibody level after SE vaccination. Alleles defined by a Lys(148)-->Met(148) polymorphism in the MHC class I alpha(2) domain were associated with spleen bacterial load after SE challenge. These results suggest that particular MHC haplotypes may contribute to control of responses to SE, and that particular polymorphisms may serve as markers for genetic resistance to SE in the chicken.