Biophysical properties of Saccharomyces cerevisiae and their relationship with HOG pathway activation

Parameterized models of biophysical and mechanical cell properties are important for predictive mathematical modeling of cellular processes. The concepts of turgor, cell wall elasticity, osmotically active volume, and intracellular osmolarity have been investigated for decades, but a consistent rigorous parameterization of these concepts is lacking. Here, we subjected several data sets of minimum volume measurements in yeast obtained after hyper-osmotic shock to a thermodynamic modeling framework. We estimated parameters for several relevant biophysical cell properties and tested alternative hypotheses about these concepts using a model discrimination approach. In accordance with previous reports, we estimated an average initial turgor of 0.6 ± 0.2 MPa and found that turgor becomes negligible at a relative volume of 93.3 ± 6.3% corresponding to an osmotic shock of 0.4 ± 0.2 Osm/l. At high stress levels (4 Osm/l), plasmolysis may occur. We found that the volumetric elastic modulus, a measure of cell wall elasticity, is 14.3 ± 10.4 MPa. Our model discrimination analysis suggests that other thermodynamic quantities affecting the intracellular water potential, for example the matrix potential, can be neglected under physiological conditions. The parameterized turgor models showed that activation of the osmosensing high osmolarity glycerol (HOG) signaling pathway correlates with turgor loss in a 1:1 relationship. This finding suggests that mechanical properties of the membrane trigger HOG pathway activation, which can be represented and quantitatively modeled by turgor

The summer bacterial and archaeal community composition of the northern Barents Sea

Climate change related alterations in the Arctic have influences on the marine ecosystems, in particular on phytoplankton bloom dynamics. Since phytoplankton blooms are the main provider of carbon sources to the microbial loop, the bacterial and archaeal community are affected by the changes as well. Warmer water and less sea ice can lead to an earlier onset of phytoplankton blooms and consequently also to changes in the bacterial and archaeal community dynamics throughout Arctic summers. Here, we compared the bacterial and archaeal community composition during three summers (2018, 2019, and 2021) along a transect from the Barents Sea to the Arctic Ocean north of Svalbard. We used 16S rRNA gene sequencing to investigate changes in the communities in time and space. The main results showed that, Gammaproteobacteria (Nitrincolaceae), Bacteroidia (Polaribacter), and Alphaproteobacteria (SAR11 clade 1a members) dominated the bacterial and archaeal community in the surface waters but varied in abundance patterns between the years. The variations are potentially a result of different phytoplankton bloom stages and consequently differences in the availability of carbon sources. The distinctly different deep water communities were dominated by Candidatus Nitrosopumilus, Marinimicrobia, and members of the SAR324 clade in all years. The results indicate that changes in phytoplankton bloom dynamics can influence bacterial and archaeal community and thereby marine carbon cycling in surface waters, although direct links to the effects of global warming remain uncertain.publishedVersio

Growth and competition in a warmer ocean: a field experiment with a non-native and two native habitat-building seaweeds

Kelps and fucoids are important members of temperate seaweed communities, but may be negatively impacted by climate change and non-native species. We used a field experiment to investigate the effect of higher temperatures and a non-native seaweed, Sargassum muticum, on the kelp Saccharina latissima and fucoid Fucus serratus. All 3 are canopy-forming species which may grow together in the infralittoral and upper sublittoral zones in southwestern Norway. Artificial assemblages with different combinations of the species were placed in the shallow sublittoral, and length changes, weight changes and survival of the thalli were measured. This was done during a hot summer and again during a cool summer. The results showed that the species and their competitive interactions were affected by the different thermal conditions. S. latissima was the most successful species in the cool summer and had an impact on the other 2 species, but it was strongly negatively affected by the hot summer. Under these conditions, F. serratus became the most successful species, gaining the most weight. The effect of Sargassum muticum on the native species was no larger than the effect of intraspecific competition within those species. At the end of both summers S. muticum was in poor condition, potentially caused by low seawater nutrients resulting in low internal nitrogen

Viruses on the menu: The appendicularian Oikopleura dioica efficiently removes viruses from seawater

Appendicularians are planktonic marine tunicates with elaborate filter-feeding houses that can efficiently trap particles as small as 0.2 lm. While marine viruses are seldom considered outside their role in disease transmission, we conducted a controlled laboratory experiment to determine if the appendicularian Oikopleura dioica can trap and ingest the Emiliania huxleyi virus (EhV; 160–180 nm diameter). Removal and retention of EhV during 2.5 h and overnight incubations at 158C were measured using flow cytometry and quantitative polymerase chain reaction specific for the mcp gene of EhV. The fate of retained EhV was tested by quantifying EhV DNA in three biological compartments: house-trapping, ingestion/digestion, and defecation. Clearance rates for EhV varied from approximately 2 mL ind21 d21 to 50 mL ind21 d21, with highest rates for 4–5 d-old animals. EhV particles were cleared by O. dioica at rates similar to those reported for larger food particles, with mean clearance rates in the 2.5 h incubations ranging from approximately 2 mL ind21 d21 to 50 mL ind21 d21. This demonstrates efficient virus removal by O. dioica and a previously overlooked link between the microbial loop and the classical marine food web. EhV DNA was readily detectable above background levels in O. dioica houses, gut contents, and faecal pellets, suggesting that appendicularian houses and faecal pellets may contribute to the dispersal of viruses. Furthermore, clearance of EhV and presumably other viruses by O. dioica may be a significant sink for viruses and thus an important factor in regulating the population dynamics of viruses and their hosts.acceptedVersio

Exploring the impact of osmoadaptation on glycolysis using time-varying response-coefficients

We present a model of osmoadaptation in S.cerevisiae based on existing experimental and theoretical work. In order to investigate the impact of osmoadaptation on glycolysis, this model focuses on the interactions between glycolysis and osmoadaptation, namely the production of glycerol and its influence on flux towards pyruvate. Evaluation of this model shows that, depending on initial relations between glycerol and pyruvate production, the increased glycerol production can have a substantial negative effect on the pyruvate production rate. Existing experimental data and a detailed analysis of the model lead to the suggestion of an interaction between activated Hog1 and activators of glycolysis such as Pfk26

Quantification of cell volume changes upon hyperosmotic stress in Saccharomyces cerevisiae.

Cell volume is a biophysical property, which is of great importance for quantitative characterisations of biological processes, such as osmotic adaptation. It also is a crucial parameter in the most common type of mathematical description of cellular behaviour-ordinary differential equation (ODE) models, e.g. the integrative model of the osmotic stress response in baker\u27s yeast (E. Klipp, B. Nordlander, R. Kruger, P. Gennemark and S. Hohmann, Nat. Biotechnol., 2005, 23, 975-982). Until recently only rough estimates of this value were available. In this study we measured the mean volume of more than 300 individual yeast cells (Saccharomyces cerevisiae). We quantitatively characterised the dependence between the relative cell volume and the concentration of osmoticum in the cell surrounding. We also followed the recovery of the cellular volume over time, as well as the influence of increased external osmolarity on the nuclear volume. We found that cell shrinkage caused by shifts in the external osmolarity is proportional to the stress intensity only up to 1000 mM NaCl. At this concentration the yeast cells shrink to approximately 55% of their unstressed volume and this volume is maintained even in the case of further osmolarity increase. We observed that returning to the initial, unstressed volume takes more than 45 minutes for stress concentrations exceeding 100 mM NaCl and that only cells treated with the latter concentration are able to fully regain their initial size within the course of the experiment. We postulate that the cytoplasm plays a protective role for the nucleus by buffering the changes in volume caused by external osmolarity shifts. In conclusion, we quantitatively characterised the dynamics of cell volume changes caused by hyperosmotic stress, providing an accurate description of a biophysical cell property, which is crucial for precise mathematical simulations of cellular processes

Hog1 phosphorylation is delayed in severe hyper-osmotic stress.

<p>Western blot of Hog1 phosphorylation in wild type treated with 400mM and 800mM NaCl at time “0”. The upper blot was treated with antibody recognizing dually phosphorylated Hog1, the lower panel with an antibody that detects total Hog1.</p