Multiallelic recognition: Nonself-dependent dimerization of the bE and bW homeodomain proteins in ustilago maydis

AbstractIn the plant pathogenic fungus Ustilago maydis, sexual and pathogenic development are controlled by the multiallelic b mating-type locus. The b locus encodes a pair of unrelated homeodomain proteins termed bE and bW, with allelic differences clustering in the N-terminal domains of both polypeptides. Only combinations of bE and bW of different allelic origin are active. We have investigated the underlying molecular mechanism for this intracellular self/nonself recognition phenomenon. By using the two-hybrid system, we were able to show that bE and bW dimerize only if they are derived from different alleles. Dimerization involves the N-terminal variable domains. Different point mutants of bE2 were isolated that function in combination with bW2. The majority of such bE2 mutant polypeptides were also able to form heterodimers with bW2 in the two-hybrid system. Nonself-dependent dimerization of bE and bW was supported with a biochemical interaction assay with immobilized proteins. Our results suggest a model for self/nonself recognition in which variable cohesive contacts direct dimerization

Isolierung und Analyse haploider Ustilago maydis Stämme, die ein b-abhängiges Expressionsmuster zeigen

Ustilago maydis ist der Erreger des Maisbeulenbrands. Vorraussetzung für eine erfolgreiche Infektion sind Fusion zweier kompatibler, haploider Zellen und die folgende Ausbildung eines dikaryotischen Filaments. Diese Prozesse werden durch die beiden Paarungstyploci a und b kontrolliert. Der a-Locus kodiert für ein biallelisches Pheromon/Pheromonrezeptor-System, das die Zell/Zell-Erkennung und die Zellfusion reguliert. Die folgende pathogene Entwicklung wird durch den multiallelischen b-Locus kontrolliert, der für zwei Homeodomänenproteine kodiert, bW und bE. Nach der Fusion der haploiden Sporidien können sich bE/bW-Heterodimere ausschließlich aus bW und bE-Proteinen unterschiedlicher Allele bilden. Diese regulieren das filamentöse Wachstum, die Penetration der Pflanzenoberfläche, das Wachstum in der Pflanze und die Tumorinduktion. Das Ziel dieser Arbeit war es, regulatorische Gene aus U.maydis zu identifizieren, die an der Kontrolle der b-abhängigen, pathogenen Entwicklung beteiligt sind. Es wurde versucht, in einem direkten Selektionsprozess haploide, pathogene Stämme zu isolieren, die aus einer REMI-Mutagenese hervorgingen. Um eine möglichst breite Mutagenese zu erreichen, wurde eine neuartige Mutagenesestrategie angewandt, die neben Geninaktivierung ("loss of function") auch eine mögliche Aktivierung der Genexpression der betroffenen Loci ("gain of function") berücksichtigte. In einer weiteren UV-Mutagenese wurden durch Nutzung von egl1 als Reportergen Stämme isoliert, die EG-Aktivität zeigten. Die Expression des b-abhängigen, aber vermutlich nicht direkt durch das bE/bW-Heterodimer regulierten Gens egl1 sollte dabei eine Mutation in einem regulatorischen Gen anzeigen, das wiederum unter der Kontrolle von b stehen könnte. Es wurde angenommen, dass die interessantesten Stämme neben egl1 weitere b-abhängige Gene exprimieren. Eine komplexe Deregulation der Genexpression b-abhängiger Gene in haploiden Zellen sollte die Zentralität des betroffenen Regulators innerhalb der b-Regulationskaskade anzeigen. Die Komplementation des Stammes MR9-1 führte zur Isolierung eines regulatorischen Gens. hda1 kodiert für ein Protein mit signifikanter Homologie zu Histondeacetylasen und ist an der Kontrolle der differentiellen Genexpression in haploiden und dikaryotischen Zellen, und später an der Sporenentwicklung im Tumor entscheidend beteiligt. Hda1 wirkt in haploiden Zellen nicht als genereller Regulator der Genexpression, sondern bestimmt ein spezifisches Set von hda1-abhängigen Genen, das sich vornehmlich aus b-abhängigen Genen und den b-Genen selbst zusammensetzt. Haploide ∆hda1-Stämme vollziehen nicht die pathogene Entwicklung; nur dikaryotische ∆hda1-Zellen führen zur Tumorentwicklung, leiten jedoch nicht die Bildung von Sporen im Tumorgewebe ein. Funktionelle und biochemische Analysen zeigen in haploiden Zellen einen hochmolekularen Hda1-Komplex, der vermutlich den Aufbau einer höher geordneten Chromatinstruktur an regulatorischen Sequenzen bestimmt. Vermutlich kann Hda1 über einen Deacetylierungsmechanismus regulatorischer Sequenzen zur Repression b-abhängiger Gene führen

Olfactory fMRI in Patients with Parkinson's Disease

Hyposmia is one of the early signs in idiopathic Parkinson's disease (PD). Olfactory stimuli were applied during fMRI scanning to show disease-related modulation of central nervous system structures and to advance our understanding of olfactory dysfunction in PD patients. All participants received either unpleasant stimuli that smelled like rotten eggs or pleasant ones that smelled like roses. Using a block design at a 1.5 T scanner we investigated a total of 8 PD patients (mean age 60 ± 10.9 years) and 13 age matched controls (mean age 58 ± 9.6 years). PD duration ranged from 1 to 9 years (mean 6.63 years); patients had an average “Unified Parkinson's Disease Rating Scale III” score of 23.25 (range, 6–46). Olfactory function was established using the “Sniffin’ Sticks” test battery. Patients tended to rate the stimuli presented during fMRI scans as less intense, but also as more pleasant than controls. fMRI results revealed differences between PD patients and controls which depended on the type of stimulation. While both pleasant and unpleasant stimulation was associated with lower activation in the amygdalo–hippocampal complex in patients compared to controls, increased activity in response to pleasant stimuli was observed in the striatum and the left inferior frontal gyrus. In contrast, unpleasant stimulation led to hypoactivation of the ventral striatum in patients (but not in controls) and did not enhance left inferior frontal activity. These results may partly reflect differences between PD patients and healthy controls in the processing of primary dimensions of odors, intensity, and valence

The Genome of Streptococcus mitis B6 - What Is a Commensal?

Streptococcus mitis is the closest relative of the major human pathogen S. pneumoniae. The 2,15 Mb sequence of the Streptococcus mitis B6 chromosome, an unusually high-level beta-lactam resistant and multiple antibiotic resistant strain, has now been determined to encode 2100 genes. The accessory genome is estimated to represent over 40%, including 75 mostly novel transposases and IS, the prophage φB6 and another seven phage related regions. Tetracycline resistance mediated by Tn5801, and an unusual and large gene cluster containing three aminoglycoside resistance determinants have not been described in other Streptococcus spp. Comparative genomic analyses including hybridization experiments on a S. mitis B6 specific microarray reveal that individual S. mitis strains are almost as distantly related to the B6 strain as S. pneumoniae. Both species share a core of over 900 genes. Most proteins described as pneumococcal virulence factors are present in S. mitis B6, but the three choline binding proteins PcpA, PspA and PspC, and three gene clusters containing the hyaluronidase gene, ply and lytA, and the capsular genes are absent in S. mitis B6 and other S. mitis as well and confirm their importance for the pathogenetic potential of S. pneumoniae. Despite the close relatedness between the two species, the S. mitis B6 genome reveals a striking X-alignment when compared with S. pneumoniae

Multiple constraints cause positive and negative feedbacks limiting grassland soil CO2efflux under CO2enrichment

Terrestrial ecosystems are increasingly enriched with resources such as atmospheric CO2that limit ecosystem processes. The consequences for ecosystem carbon cycling depend on the feedbacks from other limiting resources and plant community change, which remain poorly understood for soil CO2efflux, JCO2, a primary carbon flux from the biosphere to the atmosphere. We applied a unique CO2enrichment gradient (250 to 500 μL L-1) for eight years to grassland plant communities on soils from different landscape positions. We identified the trajectory of JCO2responses and feedbacks from other resources, plant diversity [effective species richness, exp(H)], and community change (plant species turnover). We found linear increases in JCO2on an alluvial sandy loam and a lowland clay soil, and an asymptotic increase on an upland silty clay soil. Structural equation modeling identified CO2as the dominant limitation on JCO2on the clay soil. In contrast with theory predicting limitation from a single limiting factor, the linear JCO2response on the sandy loam was reinforced by positive feedbacks from aboveground net primary productivity and exp(H), while the asymptotic JCO2response on the silty clay arose from a net negative feedback among exp(H), species turnover, and soil water potential. These findings support a multiple resource limitation view of the effects of global change drivers on grassland ecosystem carbon cycling and highlight a crucial role for positive or negative feedbacks between limiting resources and plant community structure. Incorporating these feedbacks will improve models of terrestrial carbon sequestration and ecosystem services

Multiple constraints cause positive and negative feedbacks limiting grassland soil CO2efflux under CO2enrichment

Terrestrial ecosystems are increasingly enriched with resources such as atmospheric CO2that limit ecosystem processes. The consequences for ecosystem carbon cycling depend on the feedbacks from other limiting resources and plant community change, which remain poorly understood for soil CO2efflux, JCO2, a primary carbon flux from the biosphere to the atmosphere. We applied a unique CO2enrichment gradient (250 to 500 μL L-1) for eight years to grassland plant communities on soils from different landscape positions. We identified the trajectory of JCO2responses and feedbacks from other resources, plant diversity [effective species richness, exp(H)], and community change (plant species turnover). We found linear increases in JCO2on an alluvial sandy loam and a lowland clay soil, and an asymptotic increase on an upland silty clay soil. Structural equation modeling identified CO2as the dominant limitation on JCO2on the clay soil. In contrast with theory predicting limitation from a single limiting factor, the linear JCO2response on the sandy loam was reinforced by positive feedbacks from aboveground net primary productivity and exp(H), while the asymptotic JCO2response on the silty clay arose from a net negative feedback among exp(H), species turnover, and soil water potential. These findings support a multiple resource limitation view of the effects of global change drivers on grassland ecosystem carbon cycling and highlight a crucial role for positive or negative feedbacks between limiting resources and plant community structure. Incorporating these feedbacks will improve models of terrestrial carbon sequestration and ecosystem services

CO2 enrichment and soil type additively regulate grassland productivity

The development of a predictive understanding of how atmospheric CO2 enrichment is affecting the primary productivity of the terrestrial biosphere is among the most pressing of ecological challenges. The terrestrial biosphere absorbs c. 25% of anthropogenic carbon (C) emissions (Le Quere et al., 2018). Uncertainty in CO2 effects on ecosystem C uptake is a major constraint in the prediction of C cycling and the provisioning of productivity- related ecosystem services. Grasslands cover c. 25% of the terrestrial area and are an important contributor to the global C balance (Sala et al., 1996). CO2 enrichment stimulates the aboveground net primary productivity (ANPP) of most water-limited grasslands by increasing plant water use efficiency (WUE; productivity per unit of transpiration; Morgan et al., 2004; Nowak et al., 2004; Fatichi et al., 2016), but grassland ANPP, as other ecosystem functions, is determined by drivers in addition to water availability which act simultaneously and often interactively with CO2 (Polley et al., 2011). CO2 enrichment usually shows greater stimulation of plant productivity when nitrogen (N) availability is relatively high (Owensby et al., 1994; Reich & Hobbie, 2013; Mueller et al., 2016), for example. Other drivers include precipitation timing (Hovenden et al., 2014), disturbance regimes (Newton et al., 2014), plant species composition (Langley & Megonigal, 2010; Fay et al., 2012; Polley et al., 2012) and soil properties (Epstein et al., 1997, 1998), including soil texture, which influences water availability to plants (Tor-Ngern et al., 2017)

CO2 enrichment and soil type additively regulate grassland productivity

The development of a predictive understanding of how atmospheric CO2 enrichment is affecting the primary productivity of the terrestrial biosphere is among the most pressing of ecological challenges. The terrestrial biosphere absorbs c. 25% of anthropogenic carbon (C) emissions (Le Quere et al., 2018). Uncertainty in CO2 effects on ecosystem C uptake is a major constraint in the prediction of C cycling and the provisioning of productivity- related ecosystem services. Grasslands cover c. 25% of the terrestrial area and are an important contributor to the global C balance (Sala et al., 1996). CO2 enrichment stimulates the aboveground net primary productivity (ANPP) of most water-limited grasslands by increasing plant water use efficiency (WUE; productivity per unit of transpiration; Morgan et al., 2004; Nowak et al., 2004; Fatichi et al., 2016), but grassland ANPP, as other ecosystem functions, is determined by drivers in addition to water availability which act simultaneously and often interactively with CO2 (Polley et al., 2011). CO2 enrichment usually shows greater stimulation of plant productivity when nitrogen (N) availability is relatively high (Owensby et al., 1994; Reich & Hobbie, 2013; Mueller et al., 2016), for example. Other drivers include precipitation timing (Hovenden et al., 2014), disturbance regimes (Newton et al., 2014), plant species composition (Langley & Megonigal, 2010; Fay et al., 2012; Polley et al., 2012) and soil properties (Epstein et al., 1997, 1998), including soil texture, which influences water availability to plants (Tor-Ngern et al., 2017)

Ruthenium polypyridyl complexes and their modes of interaction with DNA : is there a correlation between these interactions and the antitumor activity of the compounds?

Various interaction modes between a group of six ruthenium polypyridyl complexes and DNA have been studied using a number of spectroscopic techniques. Five mononuclear species were selected with formula [Ru(tpy) L1L2](2-n)?, and one closely related dinuclear cation of formula [{Ru(apy)(tpy)}2{l-H2N(CH2)6NH2}]4?. The ligand tpy is 2,20:60,200-terpyridine and the ligand L1 is a bidentate ligand, namely, apy (2,20-azobispyridine), 2-phenylazopyridine, or 2-phenylpyridinylmethylene amine. The ligand L2 is a labile monodentate ligand, being Cl-, H2O, or CH3CN. All six species containing a labile L2 were found to be able to coordinate to the DNA model base 9-ethylguanine by 1H NMR and mass spectrometry. The dinuclear cationic species, which has no positions available for coordination to a DNA base, was studied for comparison purposes. The interactions between a selection of four representative complexes and calf-thymus DNA were studied by circular and linear dichroism. To explore a possible relation between DNA-binding ability and toxicity, all compounds were screened for anticancer activity in a variety of cancer cell lines, showing in some cases an activity which is comparable to that of cisplatin. Comparison of the details of the compound structures, their DNA binding, and their toxicity allows the exploration of structure–activity relationships that might be used to guide optimization of the activity of agents of this class of compounds

Influence of orbital contributions to the valence band alignment of Bi2O3, Fe2O3, BiFeO3, and Bi0.5Na0.5TiO3

The formation of an interface between Bi2O3, Fe2O3, BiFeO3, Bi0.5Na0.5TiO3, and the high work function metallic RuO2 is studied using photoelectron spectroscopy with in situ RuO2 deposition. Schottky barrier heights are derived and the valence band maximum energies of the studied materials are aligned with respect to each other as well as to other functional oxides like SrTiO3 and PbTiO3. The energy band alignment follows systematic trends compared to a large number of oxides, and can be understood in terms of the contribution of Fe 3d and Bi 6s/6p (lone pair) orbitals to electronic states near the valence band maximum. The results indicate that the valence band maxima are largely determined by the local environment of the cations, which allows to estimate valence band maximum energies of oxides with multiple cations from those of their parent binary compounds. The high valence band maximum of BiFeO3 is consistent with reported p-type conduction of acceptor doped material, while the high conduction band minimum makes n-type conduction unlikely