Bacterial cell wall peptidoglycan is essential, maintaining both cellular integrity and morphology, in the face of internal turgor pressure. Peptidoglycan synthesis is important, as it is targeted by cell wall antibiotics, including methicillin and vancomycin. Here, we have used the major human pathogen Staphylococcus aureus to elucidate both the cell wall dynamic processes essential for growth (life) and the bactericidal effects of cell wall antibiotics (death) based on the principle of coordinated peptidoglycan synthesis and hydrolysis. The death of S. aureus due to depletion of the essential, two-component and positive regulatory system for peptidoglycan hydrolase activity (WalKR) is prevented by addition of otherwise bactericidal cell wall antibiotics, resulting in stasis. In contrast, cell wall antibiotics kill via the activity of peptidoglycan hydrolases in the absence of concomitant synthesis. Both methicillin and vancomycin treatment lead to the appearance of perforating holes throughout the cell wall due to peptidoglycan hydrolases. Methicillin alone also results in plasmolysis and misshapen septa with the involvement of the major peptidoglycan hydrolase Atl, a process that is inhibited by vancomycin. The bactericidal effect of vancomycin involves the peptidoglycan hydrolase SagB. In the presence of cell wall antibiotics, the inhibition of peptidoglycan hydrolase activity using the inhibitor complestatin results in reduced killing, while, conversely, the deregulation of hydrolase activity via loss of wall teichoic acids increases the death rate. For S. aureus, the independent regulation of cell wall synthesis and hydrolysis can lead to cell growth, death, or stasis, with implications for the development of new control regimes for this important pathogen.
- MeSH
- antibakteriální látky farmakologie MeSH
- antiinfekční látky metabolismus farmakologie MeSH
- bakteriální proteiny metabolismus MeSH
- buněčná stěna metabolismus fyziologie MeSH
- homeostáza MeSH
- kyseliny teichoové metabolismus MeSH
- methicilin farmakologie MeSH
- N-acetylmuramoyl-L-alaninamidasa metabolismus MeSH
- peptidoglykan metabolismus MeSH
- stafylokokové infekce mikrobiologie MeSH
- Staphylococcus aureus růst a vývoj metabolismus MeSH
- vankomycin farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Streptococcus pneumoniae is an opportunistic human pathogen that encodes a single eukaryotic-type Ser/Thr protein kinase StkP and its functional counterpart, the protein phosphatase PhpP. These signaling enzymes play critical roles in coordinating cell division and growth in pneumococci. In this study, we determined the proteome and phosphoproteome profiles of relevant mutants. Comparison of those with the wild-type provided a representative dataset of novel phosphoacceptor sites and StkP-dependent substrates. StkP phosphorylates key proteins involved in cell division and cell wall biosynthesis in both the unencapsulated laboratory strain Rx1 and the encapsulated virulent strain D39. Furthermore, we show that StkP plays an important role in triggering an adaptive response induced by a cell wall-directed antibiotic. Phosphorylation of the sensor histidine kinase WalK and downregulation of proteins of the WalRK core regulon suggest crosstalk between StkP and the WalRK two-component system. Analysis of proteomic profiles led to the identification of gene clusters regulated by catabolite control mechanisms, indicating a tight coupling of carbon metabolism and cell wall homeostasis. The imbalance of steady-state protein phosphorylation in the mutants as well as after antibiotic treatment is accompanied by an accumulation of the global Spx regulator, indicating a Spx-mediated envelope stress response. In summary, StkP relays the perceived signal of cell wall status to key cell division and regulatory proteins, controlling the cell cycle and cell wall homeostasis.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální proteiny metabolismus MeSH
- buněčná stěna účinky léků fyziologie MeSH
- fosfoproteiny metabolismus MeSH
- fosforylace MeSH
- fyziologický stres * MeSH
- proteinkinasy metabolismus MeSH
- proteom MeSH
- Streptococcus pneumoniae účinky léků fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- dataset MeSH
- práce podpořená grantem MeSH
In germinating seeds under unfavorable environmental conditions, the mobilization of stores in the cotyledons is delayed, which may result in a different modulation of carbohydrates balance and a decrease in seedling vigor. Tall fescue (Festuca arundinacea Schreb.) caryopses grown at 4°C in the dark for an extended period in complete absence of nutrients, showed an unexpected ability to survive. Seedlings grown at 4°C for 210 days were morphologically identical to seedlings grown at 23°C for 21 days. After 400 days, seedlings grown at 4°C were able to differentiate plastids to chloroplast in just few days once transferred to the light and 23°C. Tall fescue exposed to prolonged period at 4°C showed marked anatomical changes: cell wall thickening, undifferentiated plastids, more root hairs and less xylem lignification. Physiological modifications were also observed, in particular related to sugar content, GA and ABA levels and amylolytic enzymes pattern. The phytohormones profiles exhibited at 4 and 23°C were comparable when normalized to the respective physiological states. Both the onset and the completion of germination were linked to GA and ABA levels, as well as to the ratio between these two hormones. All plants showed a sharp decline in carbohydrate content, with a consequent onset of gradual sugar starvation. This explained the slowed then full arrest in growth under both treatment regimes. The analysis of amylolytic activity showed that Ca2+ played a central role in the stabilization of several isoforms. Overall, convergence of starvation and hormone signals meet in crosstalk to regulate germination, growth and development in tall fescue.
- MeSH
- alfa-amylasy metabolismus MeSH
- buněčná stěna metabolismus fyziologie MeSH
- časové faktory MeSH
- Festuca metabolismus fyziologie MeSH
- fyziologická adaptace fyziologie účinky záření MeSH
- fyziologický stres fyziologie MeSH
- gibereliny metabolismus MeSH
- kořeny rostlin metabolismus fyziologie MeSH
- kotyledon metabolismus fyziologie MeSH
- kyselina abscisová metabolismus MeSH
- lignin metabolismus MeSH
- nízká teplota MeSH
- rostlinné proteiny metabolismus MeSH
- sacharidy analýza MeSH
- semenáček fyziologie MeSH
- světlo MeSH
- tma MeSH
- vápník metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
The development and growth of plants, as well as their successful adaptation to a variety of environments, is highly dependent on the conduction of water, nutrients and other important molecules throughout the plant body. Xylem is a specialized vascular tissue that serves as a conduit of water and minerals and provides mechanical support for upright growth. Wood, also known as secondary xylem, constitutes the major part of mature woody stems and roots. In the past two decades, a number of key factors including hormones, signal transducers and (post)transcriptional regulators have been shown to control xylem formation. We outline the main mechanisms shown to be essential for xylem development in various plant species, with an emphasis on Arabidopsis thaliana, as well as several tree species where xylem has a long history of investigation. We also summarize the processes which have been shown to be instrumental during xylem maturation. This includes mechanisms of cell wall formation and cell death which collectively complete xylem cell fate.
Chromerida are photoautotrophic alveolates so far only isolated from corals in Australia. It has been shown that these secondary plastid-containing algae are closely related to apicomplexan parasites and share various morphological and molecular characters with both Apicomplexa and Dinophyta. So far, the only known representative of the phylum was Chromera velia. Here we provide a formal description of another chromerid, Vitrella brassicaformis gen. et sp. nov., complemented with a detailed study on its ultrastructure, allowing insight into its life cycle. The novel alga differs significantly from the related chromerid C. velia in life cycle, morphology as well as the plastid genome. Analysis of photosynthetic pigments on the other hand demonstrate that both chromerids lack chlorophyll c, the hallmark of phototrophic chromalveolates. Based on the relatively high divergence between C. velia and V. brassicaformis, we propose their classification into distinct families Chromeraceae and Vitrellaceae. Moreover, we predict a hidden and unexplored diversity of the chromerid algae.
- MeSH
- Alveolata klasifikace genetika izolace a purifikace fyziologie ultrastruktura MeSH
- beta-karoten fyziologie MeSH
- biologické pigmenty fyziologie MeSH
- buněčná membrána fyziologie ultrastruktura MeSH
- buněčná stěna fyziologie ultrastruktura MeSH
- chlorofyl fyziologie MeSH
- elektronová mikroskopie MeSH
- flagella fyziologie ultrastruktura MeSH
- fylogeneze MeSH
- genom plastidový MeSH
- korálové útesy MeSH
- plastidy genetika fyziologie MeSH
- spory protozoální fyziologie ultrastruktura MeSH
- xanthofyly fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Differentiation into highly resistant double-walled cysts is a major mechanism allowing amphizoic acanthamoebae to survive under long-lasting, unfavourable environmental conditions. We found that relatively low concentrations of methanol, acetone or DMSO stimulate promptAcanthamoebadifferentiation into a rounded cyst-like stage with a single envelope. To address whether this rapid response differs from the encystment, time-dependent changes in cell surface characteristics and cyst-specific gene expression were monitored in encystating cells and cells differentiating under methanol treatment using microscopic, lectin-binding, PCR and resistance studies. In contrast to the encystment: (1) a single-layered amorphous mannose/glucose coat was the only envelope assembled on the surface of the solvent-treated cells, (2) the cyst-specific protein (CSP21) was not expressed, (3) the coat did not protect cells against acidic pH and (4) in solvent-free encystment medium, the coated cells did not assemble the double-layered wall, thus indicating that these cells were not immature cysts. These findings lead us to specify a terminal stage of rapidAcanthamoebadifferentiation elicited by acute organic solvent stress as "pseudocyst", and to suggest that encystation and pseudocyst formation are distinct stress responses. Moreover, the possibility exists that pseudocysts might form in response to certain contact lens solutions thus increasing resistance of acanthamoebae to disinfecting agents.
- MeSH
- Acanthamoeba účinky léků fyziologie ultrastruktura MeSH
- buněčná stěna fyziologie ultrastruktura MeSH
- dimethylsulfoxid farmakologie MeSH
- ethanol farmakologie MeSH
- fyziologická adaptace MeSH
- fyziologický stres MeSH
- genotyp MeSH
- methanol farmakologie MeSH
- rozpouštědla MeSH
- stanovení celkové genové exprese MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Vlastnosti povrchu jsou pro jednobuněčné organismy velmi důležité, protože rozhraní mezi buněčnými obaly a vnějším prostředím hraje významnou roli v celkové fyziologii buňky. Vnější povrch buňky zprostředkovává výměnné a adhezivní procesy, účastní se růstu buněk i buněčného dělení a ovlivňuje interakce s imunologickými faktory. Znalosti o povrchových vlastnostech buněk rozšiřují nejen možnost pochopení fyziologie buněk, ale přináší i informace využitelné v biotechnologiích a medicíně. Tento článek shrnuje nejčastěji používané metody pro zkoumání fyzikálně-chemických vlastností povrhu buněk.
The surface properties are very importent for unicellular organisms, because interface between cell envelope and outer environmet plays an important role in overall physiology of cells. The outer cell surface mediates exchange and adhesive processes, participates in cell growth and division and also influences interactions with immunological factors. Understanding the cell surface properties can shed light on cell fysiology and also bring information usefull for biotechnology and medicine. This article summarize the most frequently used methods for studying physico-chemical properties of cell surface.
