Biology of archaea from a novel family Cuniculiplasmataceae (Thermoplasmata) ubiquitous in hyperacidic environments

The order Thermoplasmatales (Euryarchaeota) is represented by the most acidophilic organisms known so far that are poorly amenable to cultivation. Earlier culture-independent studies in Iron Mountain (California) pointed at an abundant archaeal group, dubbed ‘G-plasma’. We examined the genomes and physiology of two cultured representatives of a Family Cuniculiplasmataceae, recently isolated from acidic (pH 1–1.5) sites in Spain and UK that are 16S rRNA gene sequence-identical with ‘G-plasma’. Organisms had largest genomes among Thermoplasmatales (1.87–1.94 Mbp), that shared 98.7–98.8% average nucleotide identities between themselves and ‘G-plasma’ and exhibited a high genome conservation even within their genomic islands, despite their remote geographical localisations. Facultatively anaerobic heterotrophs, they possess an ancestral form of A-type terminal oxygen reductase from a distinct parental clade. The lack of complete pathways for biosynthesis of histidine, valine, leucine, isoleucine, lysine and proline pre-determines the reliance on external sources of amino acids and hence the lifestyle of these organisms as scavengers of proteinaceous compounds from surrounding microbial community members. In contrast to earlier metagenomics-based assumptions, isolates were S-layer-deficient, non-motile, non-methylotrophic and devoid of iron-oxidation despite the abundance of methylotrophy substrates and ferrous iron in situ, which underlines the essentiality of experimental validation of bioinformatic predictions

Life-cycle analysis of coesite-bearing garnet

Detrital coesite-bearing garnet is the final product of a complex geological cycle including coesite entrapment at ultrahigh-pressure conditions, exhumation to Earth’s surface, erosion, and sedimentary transport. In contrast to the usual enrichment of high-grade metamorphic garnet in 14 medium- to coarse-sand fractions, coesite-bearing grains are often enriched in the very fine-sand fraction. To understand this imbalance, we analyze the role of source rock lithology, inclusion size, inclusion frequency, and fluid infiltration on the grain-size heterogeneity of coesite-bearing garnet based on a dataset of 2100 inclusion-bearing grains, of which 93 contain coesite, from the Saxonian Erzgebirge, Germany. By combining inclusion assemblages and garnet chemistry, we show that mafic garnet contains a low number of coesite inclusions per grain and is enriched in the coarse fraction, and felsic garnet contains variable amounts of coesite inclusions per grain, whereby coesite-poor grains are enriched in the coarse fraction and coesite-rich grains extensively disintegrated into smaller fragments resulting in an enrichment in the fine fraction. Raman images reveal that small coesite inclusions <9 µm are primarily monomineralic, whereas larger inclusions partially transformed to quartz, and garnet fracturing, fluid infiltration, and the coesite-to-quartz transformation is a late process during exhumation taking place at ~330°C. A model for the disintegration of coesite-bearing garnet enables explaining the heterogeneous grain27 size distribution by inclusion frequency. High abundances of coesite inclusions cause a high degree of fracturing and fracture connections to smaller inclusions, allowing fluid infiltration and the transformation to quartz, which in turn further promotes garnet disintegration

Bacterial microcompartment-directed polyphosphate kinase promotes stable polyphosphate accumulation in E. coli

Processes for the biological removal of phosphate from wastewater rely on temporary manipula-tion of bacterial polyphosphate levels by phased environmental stimuli. In E. coli polyphosphate levels are controlled via the polyphosphate-synthesizing enzyme polyphosphate kinase (PPK1) and exopolyphosphatases (PPX and GPPA), and are temporarily enhanced by PPK1 overexpression and reduced by PPX overexpression. We hypothesized that partitioning PPK1 from cytoplasmic exopoly phosphatases would increase and stabilize E. coli polyphosphate levels. Partitioning was achieved by co-expression of E. coli PPK1 fused with a microcompartment-targeting sequence and an artificial operon of Citrobacter freundii bacterial microcompartment genes. Encapsulation of targeted PPK1 resulted in persistent phosphate uptake and stably increased cellular polyphos-phate levels throughout cell growth and into the stationary phase, while PPK1 overexpression alone produced temporary polyphosphate increase and phosphate uptake. Targeted PPK1 increased polyphosphate in microcompartments eight-fold compared with non-targeted PPK1. Co-expression of PPX polyphosphatase with targeted PPK1 had little effect on elevated cellular polyphosphate levels because microcompartments retained polyphosphate. Co-expression of PPX with non-targeted PPK1 reduced cellular polyphosphate levels. Thus, subcellular compartmentali-zation of a polymerizing enzyme sequesters metabolic products from competing catabolism by preventing catabolic enzyme access. Specific application of this process to polyphosphate is of potential application for biological phosphate removal

Detrital garnet petrology challenges Paleoproterozoic ultrahigh-pressure metamorphism in western Greenland

Modern-style plate tectonics is characterised by the global operation of cold and deep subduction involving blueschist facies and ultrahigh-pressure metamorphism. This has been a common process since the Neoproterozoic, but a couple of studies indicate similar processes were active in the Paleoproterozoic, at least on the local scale. Particularly conspicuous are extreme ultrahigh-pressure conditions of ∼ 7 GPa at thermal gradients &lt; 150 ∘C GPa−1 proposed for metamorphic rocks of the Nordre Strømfjord shear zone in the western part of the Paleoproterozoic Nagssugtoqidian Orogen of Greenland. By acquiring a large dataset of heavy minerals (n = 52 130) and garnet major-element composition integrated with mineral inclusion analysis (n=2669) from modern sands representing fresh and naturally mixed erosional material from the metamorphic rocks, we here intensely screened the area for potential occurrences of ultrahigh-pressure rocks and put constraints on the metamorphic evolution. Apart from the absence of any indications pointing to ultrahigh-pressure and low-temperature–high-pressure metamorphism, the results are well in accordance with a common Paleoproterozoic subduction–collision metamorphic evolution along a Barrovian-type intermediate temperature and pressure gradient with a pressure peak at the amphibolite–granulite–eclogite-facies transition and a temperature peak at medium- to high-pressure granulite-facies conditions. In addition, we discuss that all “evidence” for ultrahigh-pressure metamorphism proposed in the literature for rocks of this area is equivocal. Accordingly, the Nordre Strømfjord shear zone is not an example of modern-style plate tectonics in the Paleoproterozoic or of very low thermal gradients and extreme pressure conditions in general.</p

An Angiotensin I-Converting Enzyme Mutation (Y465D) Causes a Dramatic Increase in Blood ACE via Accelerated ACE Shedding

Angiotensin I-converting enzyme (ACE) metabolizes a range of peptidic substrates and plays a key role in blood pressure regulation and vascular remodeling. Thus, elevated ACE levels may be associated with an increased risk for different cardiovascular or respiratory diseases. Previously, a striking familial elevation in blood ACE was explained by mutations in the ACE juxtamembrane region that enhanced the cleavage-secretion process. Recently, we found a family whose affected members had a 6-fold increase in blood ACE and a Tyr465Asp (Y465D) substitution, distal to the stalk region, in the N domain of ACE.HEK and CHO cells expressing mutant (Tyr465Asp) ACE demonstrate a 3- and 8-fold increase, respectively, in the rate of ACE shedding compared to wild-type ACE. Conformational fingerprinting of mutant ACE demonstrated dramatic changes in ACE conformation in several different epitopes of ACE. Cell ELISA carried out on CHO-ACE cells also demonstrated significant changes in local ACE conformation, particularly proximal to the stalk region. However, the cleavage site of the mutant ACE--between Arg1203 and Ser1204--was the same as that of WT ACE. The Y465D substitution is localized in the interface of the N-domain dimer (from the crystal structure) and abolishes a hydrogen bond between Tyr465 in one monomer and Asp462 in another.The Y465D substitution results in dramatic increase in the rate of ACE shedding and is associated with significant local conformational changes in ACE. These changes could result in increased ACE dimerization and accessibility of the stalk region or the entire sACE, thus increasing the rate of cleavage by the putative ACE secretase (sheddase)

The Photosynthetic Apparatus and Its Regulation in the Aerobic Gammaproteobacterium Congregibacter litoralis gen. nov., sp. nov

BACKGROUND: There is accumulating evidence that in some marine environments aerobic bacteriochlorophyll a-producing bacteria represent a significant part of the microbial population. The interaction of photosynthesis and carbon metabolism in these interesting bacteria is still largely unknown and requires further investigation in order to estimate their contribution to the marine carbon cycle. METHODOLOGY/PRINCIPAL FINDINGS: Here, we analyzed the structure, composition and regulation of the photosynthetic apparatus in the obligately aerobic marine gammaproteobacterium KT71(T). Photoheterotrophically grown cells were characterized by a poorly developed lamellar intracytoplasmic membrane system, a type 1 light-harvesting antenna complex and a photosynthetic reaction center associated with a tetraheme cytochrome c. The only photosynthetic pigments produced were bacteriochlorophyll a and spirilloxanthin. Under semiaerobic conditions KT71(T) cells expressing a photosynthetic apparatus showed a light-dependent increase of growth yield in the range of 1.3-2.5 fold. The expression level of the photosynthetic apparatus depended largely on the utilized substrate, the intermediary carbon metabolism and oxygen tension. In addition, pigment synthesis was strongly influenced by light, with blue light exerting the most significant effect, implicating that proteins containing a BLUF domain may be involved in regulation of the photosynthetic apparatus. Several phenotypic traits in KT71(T) could be identified that correlated with the assumed redox state of growing cells and thus could be used to monitor the cellular redox state under various incubation conditions. CONCLUSIONS/SIGNIFICANCE: In a hypothetical model that explains the regulation of the photosynthetic apparatus in strain KT71(T) we propose that the expression of photosynthesis genes depends on the cellular redox state and is maximal under conditions that allow a balanced membrane redox state. So far, bacteria capable of an obligately aerobic, photosynthetic metabolism constitute a unique phenotype within the class Gammaproteobacteria, so that it is justified to propose a new genus and species, Congregibacter litoralis gen. nov, sp. nov., represented by the type strain KT71(T) ( = DSM 17192(T) = NBRC 104960(T))

Exopolymer production and microcolony formation by planktonic freshwater bacteria: defence against protistan grazing

The defence mechanisms of 2 novel bacterial isolates against protistan grazing were investigated in experiments with batch and continuous cultures. Strains MWH55 and MWH73 were isolated from the plankton of mesotrophic lakes using culture conditions with strong flagellate grazing pressure. The analysis of their 16S rRNA genes revealed that both strains belong to the Betaproteobacteria and demonstrated a close phylogenetic relatedness to bacteria previously detected by culture-independent methods in river biofilms and lake snow aggregates. Both strains showed a very weak sensitivity to flagellate predation, and both formed planktonic microcolonies that exceeded the upper size limit for ingestion by flagellates, and were thus protected from predation. These microcolonies consisted of cells embedded in a common exopolymeric matrix, yet lacked direct cell-to-cell contact between neighbouring cells. The structure of the exopolymeric matrix of the microcolonies was visualized by electron microscopy, while detection of exopolysaccharides by several fluorescently labeled lectins failed. The matrix possessed a complex 3-dimensional structure with strain-specific characteristics. Currently, it is not clear whether the 2 isolates possess a planktonic life strategy or whether they switch between sessile (biofilm or lake snow aggregates) and planktonic dispersal stages. In both cases, however, a low sensitivity of the planktonic stages to protistan predation is ecologically advantageous. The ability to form an exopolymeric matrix enabling the formation of predation-protected microcolonies is assumed to play a key role in the ecology of these bacteria