The structural characterization of a prophage-encoded extracellular DNase from Streptococcus pyogenes

The pathogenic bacterium Group A Streptococcus pyogenes produces several extracellular DNases that have been shown to facilitate invasive infection by evading the human host immune system. DNases degrade the chromatin in neutrophil extracellular traps, enabling the bacterium to evade neutrophil capture. Spd1 is a type I, nonspecific ββα/metal-dependent nuclease from Streptococcus pyogenes, which is encoded by the SF370.1 prophage and is likely to be expressed as a result of prophage induction. We present here the X-ray structure of this DNase in the wild-type and Asn145Ala mutant form. Through structural and sequence alignments as well as mutagenesis studies, we have identified the key residues His121, Asn145 and Glu164, which are crucial for Spd1 nucleolytic activity and shown the active site constellation. Our wild-type structure alludes to the possibility of a catalytically blocked dimeric form of the protein. We have investigated the multimeric nature of Spd1 using size-exclusion chromatography with multi-angle light scattering (SEC-MALLS) in the presence and absence of the divalent metal ion Mg2+, which suggests that Spd1 exists in a monomeric form in solution

Structural and mechanistic insights into a Bacteroides vulgatus retaining N-acetyl-β-galactosaminidase that uses neighbouring group participation

Bacteroides vulgatus is a member of the human microbiota whose abundance is increased in patients with Crohn's disease. We show that a B. vulgatus glycoside hydrolase from the carbohydrate active enzyme family GH123, BvGH123, is an N-acetyl-β-galactosaminidase that acts with retention of stereochemistry, and, through a 3-D structure in complex with Gal-thiazoline, provide evidence in support of a neighbouring group participation mechanism

Structure Guided Design and Synthesis of a Pyridazinone Series of Trypanosoma cruzi Proteasome Inhibitors

There is an urgent need for new treatments for Chagas disease, a parasitic infection which mostly impacts South and Central America. We previously reported on the discovery of GSK3494245/DDD01305143, a preclinical candidate for visceral leishmaniasis which acted through inhibition of the Leishmania proteasome. A related analogue, active against Trypanosoma cruzi, showed suboptimal efficacy in an animal model of Chagas disease, so alternative proteasome inhibitors were investigated. Screening a library of phenotypically active analogues against the T. cruzi proteasome identified an active, selective pyridazinone, the development of which is described herein. We obtained a cryo-EM co-structure of proteasome and a key inhibitor and used this to drive optimization of the compounds. Alongside this, optimization of the absorption, distribution, metabolism, and excretion (ADME) properties afforded a suitable compound for mouse efficacy studies. The outcome of these studies is discussed, alongside future plans to further understand the series and its potential to deliver a new treatment for Chagas disease.</p

Central Crosstalk for Somatic Tinnitus: Abnormal Vergence Eye Movements

Frequent oulomotricity problems with orthoptic testing were reported in patients with tinnitus. This study examines with objective recordings vergence eye movements in patients with somatic tinnitus patients with ability to modify their subjective tinnitus percept by various movements, such as jaw, neck, eye movements or skin pressure.Vergence eye movements were recorded with the Eyelink II video system in 15 (23–63 years) control adults and 19 (36–62 years) subjects with somatic tinnitus.1) Accuracy of divergence but not of convergence was lower in subjects with somatic tinnitus than in control subjects. 2) Vergence duration was longer and peak velocity was lower in subjects with somatic tinnitus than in control subjects. 3) The number of embedded saccades and the amplitude of saccades coinciding with the peak velocity of vergence were higher for tinnitus subjects. Yet, saccades did not increase peak velocity of vergence for tinnitus subjects, but they did so for controls. 4) In contrast, there was no significant difference of vergence latency between these two groups.The results suggest dysfunction of vergence areas involving cortical-brainstem-cerebellar circuits. We hypothesize that central auditory dysfunction related to tinnitus percept could trigger mild cerebellar-brainstem dysfunction or that tinnitus and vergence dysfunction could both be manifestations of mild cortical-brainstem-cerebellar syndrome reflecting abnormal cross-modality interactions between vergence eye movements and auditory signals

Characterisation of SEQ0694 (PrsA/PrtM) of Streptococcus equi as a functional peptidyl-prolyl isomerase affecting multiple secreted protein substrates

YesPeptidyl-prolyl isomerase (PPIase) lipoproteins have been shown to influence the virulence of a number of Gram-positive bacterial human and animal pathogens, most likely through facilitating the folding of cell envelope and secreted virulence factors. Here, we used a proteomic approach to demonstrate that the Streptococcus equi PPIase SEQ0694 alters the production of multiple secreted proteins, including at least two putative virulence factors (FNE and IdeE2). We demonstrate also that, despite some unusual sequence features, recombinant SEQ0694 and its central parvulin domain are functional PPIases. These data add to our knowledge of the mechanisms by which lipoprotein PPIases contribute to the virulence of streptococcal pathogens

Can the black bog ant (Formica picea Nyl.) survive in the Bieszczady National Park (SE Poland)

In the peat bogs of the Bieszczady National Park an inventory of the Black bog ant (Formica picea Nyl.) was made in order to get an impression of the probability of its survival in SE Poland. Three habitat networks are distinguished, which are situated too far from each other for (re)colonization by means of flying queens. The largest network includes five peat bogs. In two of them F. picea was found. The species was not found in peat bogs which belong to the other habitat networks. The quality of the peat bogs depends on management strategies, currently challenged by two main threats: desiccation and the influx of nutrients. This process will speed up the succession of the vegetation. As a result dwarf shrubs are encroaching on habitat areas of F. picea, where Sphagnum species are still dominating in the moss laye

Dispersal for survival: some observations on the trunk ant (Formica truncorum Fabricius)

The survival chance of the trunk ant (Formica truncorum) is compared with the survival chance of two other species of red wood ants: F.rufa and F.polyctena. Nest populations of F.truncorum are much smaller than nest populations of the other red wood ant species, which makes the species a weaker competitor. Moreover, F.truncorum has to move more often, as its nests are small and untidily built, which makes the species more dependent on external heat for raising its brood. In order to survive, the greater extinction chance of F.truncorum should be compensated by a greater colonization chance. The colonization chance depends on the number of queens produced, their chance of becoming fertilized and their chance of becoming adopted by F.fusca. It appeared that nest populations of F. truncorum produced less sexuals than those of the other red wood ant species, while there are no good arguments to assume that their queens will have a greater chance of becoming fertilized or that the queens are more skillful in finding a suitable nesting place. It is argued that local populations of the trunk ant are more prone to extinction than those of F.polyctena and F.rufa, at least in the sandy areas of west and central Europe, where most F.truncorum colonies are monodomous and largely also monogynous. So F.truncorum queens should have a greater chance to become adopted by F. fusca than F. rufa and F.polyctena. Adoption experiments can solve this problem

Genetic polymorphism in "mixed" colonies of wood ants (Hymenoptera: Formicidae) in southern Finland and its possible origin

Wood ant colonies that appear to consist of individuals representing different species are described in several previous papers. The present study is the first to elucidate the genetic basis of the spectacular morphological variability observed within such colonies. Two seemingly mixed colonies (FM-1 and FM-2) from southern Finland were investigated. On the basis of the morphology of their workers these colonies were comprised of individuals with phenotypes typical of Formica rufa L., F. polyctena Först., and F. aquilonia Yarr. The sequence of an mtDNA fragment (5' end of the cytochrome b gene) was used to examine the phylogenetic relationships among haplotypes of workers from homogeneous colonies of different wood ant species and the two supposedly mixed colonies, and to sort the individuals within the colonies into matrilines. Six microsatellite loci were used to analyse the genetic differentiation between colonies and among workers within colonies, and to detect putative hybrids. The results show that, independently of their phenotype, workers from the "mixed" colonies were genetically more similar to other individuals in their colony than to those in the homogeneous F. rufa, F. polyctena or F. aquilonia colonies. However, while colony FM-1 consisted of offspring of the same queen or more likely several maternally related queens, colony FM-2 consisted of the offspring of at least four unrelated queens. The data suggest hybridisation between F. polyctena × F. aquilonia and F. polyctena × F. rufa (and possibly subsequent mating between these hybrids) as the most probable mechanism leading to the existence of these two colonies, which implies that the hybrids are fertile. This study shows that colonies of wood ant hybrids can arise spontaneously and persist under natural conditions. The results also revealed that even some morphologically homogeneous colonies are genetically heterogeneous. In the case of closely related, morphologically similar species that interbreed, morphology can be a bad predictor of genetic differences between individuals