The complex for replication initiation of<Emphasis Type="Italic">Escherichia coli</Emphasis>

We probed the complex betweenoriC and DnaA protein using two types of mutants inoriC. Base changes in the DnaA binding sites, DnaA boxes, had little effect on origin function. Mutations which change the distance between DnaA boxes R3 and R4, on the other hand, inactivatedoriC unless the mutation deleted or inserted one complete helical turn. Origins with other 10 base pair insertions in the interval between DnaA boxes R2 and R3 were functional, but not insertions in the R1–R2 interval. FIS protein binds to a bipartite site inoriC between DnaA boxes R2 and R3. A model for theoriC/DnaA complex based on these results suggests an array of DnaA monomers with a 34 Å spacing upon whichoriC is arranged.     

Identifying and evaluating causes of alternative community states in wetland plant communities

Ecological communities can vary greatly in species composition. Often this variation is discontinuous, in that abrupt changes in composition occur over small distances in space or short periods of time. A wide range of hypotheses from different subfields of ecology have been proposed to explain these patterns. I suggest a framework to quantitatively evaluate these hypotheses with observational data by characterizing 1) how community composition varies across sites in space, 2) how community composition varies through time, and 3) the possible drivers of this variation. I applied this approach to understand the community composition of producers in temporary and semipermanent wetlands in Michigan, USA. I identified several distinct community states which were variously dominated by particular plant functional groups (submerged, floating or emergent plants) or had no plants throughout a season. Evaluating possible hypotheses to explain this variation, I found that similar communities were not necessarily clustered near each other, suggesting that dispersal was not limited for these plants. Some sites exhibited a great deal of change in plant composition among years, shifting between two community states, but there was relatively little change at sites within a year. Moreover, these shifts did not occur in a particular order to suggest directional change or repeating cycles. Community composition was associated with several environmental variables such as pH, light and depth, and multivariate analyses suggested that species had complex, nonlinear responses to these possible drivers. Alternative stable states and interactions among multiple nonlinear drivers best explained the patterns observed in these wetlands. By formalizing initial data collection in other systems with the framework suggested here, we may gain insight into the causes of alternative community states beyond wetlands and the role of climate change and other anthropogenic forces in precipitating transitions between states.     

Influence of ecological conditions on magnesium and strontium content and ultrastructure of the calcareous skeleton ofBornetella sphaerica (Zarnadini), Dasycladales

The influence of temperature and light intensity on the ultrastructure of the calcareous skeleton of theBornetella sphaerica (Dasycladales) as well as on the incorporation of Mg and Sr into the skeleton was studied. Cells were grown in laboratory culture under various light and temperature conditions in a medium which allows them to calcify as they do in their natural habitat. The results are compared with general observations and measurements reported in the literature, especially with data on algae.     

Dr. Dr. h. c. Kurt Lindner zur Vollendung des 70. Lebensjahres

European Journal of Wildlife Research -     

Lot6p from Saccharomyces cerevisiae is a FMN-dependent reductase with a potential role in quinone detoxification

NAD(P)H:quinone acceptor oxidoreductases are flavoenzymes expressed in the cytoplasm of many tissues and afford protection against the cytotoxic effects of electrophilic quinones by catalyzing a strict two-electron reduction. Such enzymes have been reported from several mammalian sources, e.g. human, mouse and rat, and from plant species. Here, we report identification of Lot6p (YLR011wp), the first soluble quinone reductase from the unicellular model organism Saccharomyces cerevisiae. Localization studies using an antibody raised against Lot6p as well as microscopic inspection of Lot6p-GFP demonstrated accumulation of the enzyme in the cytosol of yeast cells. Despite sharing only 23% similarity to type 1 human quinone reductase, Lot6p possesses biochemical properties that are similar to its human counterpart. The enzyme catalyzes a two-electron reduction of a series of natural and artificial quinone substrates at the expense of either NADH or NADPH. The kinetic mechanism follows a ping-pong bi-bi reaction scheme, with K(M) values of 1.6-11 microm for various quinones. Dicoumarol and Cibacron Marine, two well-known inhibitors of the quinone reductase family, bind to Lot6p and inhibit its activity. In vivo experiments demonstrate that the enzymatic activity of Lot6p is consistent with the phenotype of both Deltalot6 and Lot6p overexpressing strains, suggesting that Lot6p may play a role in managing oxidative stress in yeast.     

CpG and LPS can interfere negatively with prion clearance in macrophage and microglial cells

Cells of the innate immune system play important roles in the progression of prion disease after peripheral infection. It has been found in vivo and in vitro that the expression of the cellular prion protein (PrP(c)) is up-regulated on stimulation of immune cells, also indicating the functional importance of PrP(c) in the immune system. The aim of our study was to investigate the impact of cytosine-phosphate-guanosine- and lipopolysaccharide-induced PrP(c) up-regulation on the uptake and processing of the pathological prion protein (PrP(Sc)) in phagocytic innate immune cells. For this purpose, we challenged the macrophage cell line J774, the microglial cell line BV-2 and primary bone marrow-derived macrophages in a resting or stimulated state with various prion strains, and monitored the uptake and clearance of PrP(Sc). Interestingly, stimulation led either to a transient increase in the level of PrP(Sc) relative to unstimulated cells or to a decelerated degradation of PrP(Sc). These features were dependent on cell type and prion strain. Our data indicate that the stimulation of innate immune cells may be able to support transient prion propagation, possibly explained by an increased PrP(c) cell surface expression in stimulated cells. We suggest that stimulation of innate immune cells can lead to an imbalance between the propagation and degradation of PrP(Sc).