N-Terminal Domain of Nuclear IL-1α Shows Structural Similarity to the C-Terminal Domain of Snf1 and Binds to the HAT/Core Module of the SAGA Complex

Interleukin-1α (IL-1α) is a proinflammatory cytokine and a key player in host immune responses in higher eukaryotes. IL-1α has pleiotropic effects on a wide range of cell types, and it has been extensively studied for its ability to contribute to various autoimmune and inflammation-linked disorders, including rheumatoid arthritis, Alzheimer's disease, systemic sclerosis and cardiovascular disorders. Interestingly, a significant proportion of IL-1α is translocated to the cell nucleus, in which it interacts with histone acetyltransferase complexes. Despite the importance of IL-1α, little is known regarding its binding targets and functions in the nucleus. We took advantage of the histone acetyltransferase (HAT) complexes being evolutionarily conserved from yeast to humans and the yeast SAGA complex serving as an epitome of the eukaryotic HAT complexes. Using gene knock-out technique and co-immunoprecipitation of the IL-1α precursor with TAP-tagged subunits of the yeast HAT complexes, we mapped the IL-1α-binding site to the HAT/Core module of the SAGA complex. We also predicted the 3-D structure of the IL-1α N-terminal domain, and by employing structure similarity searches, we found a similar structure in the C-terminal regulatory region of the catalytic subunit of the AMP-activated/Snf1 protein kinases, which interact with HAT complexes both in mammals and yeast, respectively. This finding is further supported with the ability of the IL-1α precursor to partially rescue growth defects of snf1Δ yeast strains on media containing 3-Amino-1,2,4-triazole (3-AT), a competitive inhibitor of His3. Finally, the careful evaluation of our data together with other published data in the field allows us to hypothesize a new function for the ADA complex in SAGA complex assembly.     

Calcium Influx Rescues Adenylate Cyclase-Hemolysin from Rapid Cell Membrane Removal and Enables Phagocyte Permeabilization by Toxin Pores

Bordetella adenylate cyclase toxin-hemolysin (CyaA) penetrates the cytoplasmic membrane of phagocytes and employs two distinct conformers to exert its multiple activities. One conformer forms cation-selective pores that permeabilize phagocyte membrane for efflux of cytosolic potassium. The other conformer conducts extracellular calcium ions across cytoplasmic membrane of cells, relocates into lipid rafts, translocates the adenylate cyclase enzyme (AC) domain into cells and converts cytosolic ATP to cAMP. We show that the calcium-conducting activity of CyaA controls the path and kinetics of endocytic removal of toxin pores from phagocyte membrane. The enzymatically inactive but calcium-conducting CyaA-AC⁻ toxoid was endocytosed via a clathrin-dependent pathway. In contrast, a doubly mutated (E570K+E581P) toxoid, unable to conduct Ca²⁺ into cells, was rapidly internalized by membrane macropinocytosis, unless rescued by Ca²⁺ influx promoted in trans by ionomycin or intact toxoid. Moreover, a fully pore-forming CyaA-ΔAC hemolysin failed to permeabilize phagocytes, unless endocytic removal of its pores from cell membrane was decelerated through Ca²⁺ influx promoted by molecules locked in a Ca²⁺-conducting conformation by the 3D1 antibody. Inhibition of endocytosis also enabled the native B. pertussis-produced CyaA to induce lysis of J774A.1 macrophages at concentrations starting from 100 ng/ml. Hence, by mediating calcium influx into cells, the translocating conformer of CyaA controls the removal of bystander toxin pores from phagocyte membrane. This triggers a positive feedback loop of exacerbated cell permeabilization, where the efflux of cellular potassium yields further decreased toxin pore removal from cell membrane and this further enhances cell permeabilization and potassium efflux.     

Spatial variation in vegetation and abiotic factors related to the occurrence of a ring‐forming sedge

Abstract. The clonal sedge Carex humilis forms rings of densely aggregated ramets in a dry grassland community in Central Europe. We describe the small‐scale spatial variation, both in abiotic factors and vegetation, in relation to these rings. Compared to the surrounding vegetation the cover of plants, other than C. humilis, was significantly lower both in the central area of rings and within the rings themselves. The vegetation structure was also different. The soil was more fertile in the central area and within the ring than in the surroundings, measured both directly and by the abiotic response values of the vascular plants. We conclude that neither resource depletion nor competition from other plants were likely to be responsible for the low ramet density in the central area of C. humilis rings. Instead, we suggest that the ring form is caused either by the deposition of growth inhibiting substances or by intrinsic morphological rules.     

Vegetation on anthropogenic metalliferous soils in the Eastern Alps

A syntaxonomic revision of vegetation of anthropogenic metalliferous habitats (mine spoils of lead, zinc and copper mining) in the Eastern Alps (Austria, Germany, Italy and Slovenia) was made. The communities studied belong to theThlaspietea rotundifolii (Linaria alpina-Cerastium uniflorum comm.,Papaveri kerneri-Thlaspietum kerneri, Violetum dubyanae, Thlaspietum cepaeifolii, Thlaspietum rotundifolii, Scrophulario juratensis-Erysimetum sylvestris, Minuartia gerardii-Silene glareosa comm.,Epipactido atrorubentis-Silenetum glareosae andSileno alpestris-Moehringietum muscosae) and to theAsplenietea trichomanis (Sileno rupestris-Asplenietum). There is neither floristic support nor syntaxonomic justification for the concepts of theGalio anisophylli-Minuartion vernae, theVioletalia calaminariae and theVioletea calaminariae in the Eastern Alps. These units should be included within theThlaspion rotundifolii, Thlaspietalia rotundifolii andThlaspietea rotundifolii, respectively.     

The high-rank syntaxa of the rock-cliff and scree vegetation of the mainland Greece and Crete

A revision was undertaken of the high-rank syntaxa of the vegetation occurring in rock fissures, ledges and screes of mainland Greece and Crete. All published phytosociological relevés available were collected and subjected to numerical classification and ordination. Four orders (Androsacetalia vandelii, Onosmetalia frutescentis, Potentilletalia speciosae andPetromaruletalia pinnatae) comprising 8 alliances (one of them new) were distinguished within theAsplenietea trichomanis (rock fissures, clefts, and ledges). The scree vegetation was classified partly within theThlaspietea rotundifolii (Drypidetalia spinosae with 2 alliances) and partly within theDaphno-Festucetea (Saturejo-Scutellarietalia—a new order, with 2 alliances). Ordination revealed clear differences in floristic composition of the alliances distinguished. The studied chasmophytic vegetation of Greece contains a very high proportion of endemics which serve as diagnostic species of the syntaxa. Further, an analysis of chorological species spectra suggested that the chorological homogeneity of a syntaxon should be considered an important diagnostic feature at high-syntaxon levels.     

Different binding sites for glucose and sorbose at the erythrocyte membrane,studied by gel filtration and infrared spectroscopy

Summary Human red blood cell membranes were solubilized with sodium dodecylsulfate and incubated with various concentrations of¹⁴C-glucose and¹⁴C-sorbose. After gel filtration on Sephadex G-100, which separated lipoproteins of differing lipid content, it was observed that the radioactivity of the bound glucose coincided with the protein peak. Radioactivity of bound sorbose was found mainly before and after the protein peak. This distribution of bound sugars was confirmed by double labeling experiments in which³H-glucose and¹⁴C-sorbose were applied simultaneously. Infrared spectroscopy revealed differences between the membranes loaded with sorbose and glucose. Particularly, the band in the C–O–C and P=O region at 1,225 cm^–1 was intensified in the sorbose-loaded membranes. Compared to serum albumin, the erythrocyte membranes were found to bind 4 times as much¹⁴C-glucose per mg of protein. It is concluded from the results obtained by gel filtration that glucose and sorbose preferentially bind at different sites of the erythrocyte membrane. The results obtained by infrared spectroscopy correspond with this conclusion.