scholarly journals Teichoic acids anchor distinct cell wall lamellae in an apically growing bacterium

2019 ◽  
Author(s):  
Eveline Ultee ◽  
Lizah T. van der Aart ◽  
Dino van Dissel ◽  
Christoph A. Diebolder ◽  
Gilles P. van Wezel ◽  
...  

AbstractThe bacterial cell wall is a dynamic, multicomponent structure that provides structural support for cell shape and physical protection from the environment. In monoderm species, the thick cell wall is made up predominantly of peptidoglycan, teichoic acids and a variety of capsular glycans. Filamentous monoderm Actinobacteria, such as Streptomyces coelicolor, incorporate new cell wall material at the apex of their hyphal cells during growth. In this study we use cryo-electron tomography to reveal the structural architecture of the cell wall of this bacterium. Our data shows a density difference between the apex and subapical regions of chemically isolated sacculi. Removal of the teichoic acids with hydrofluoric acid reveals a rough and patchy cell wall and distinct lamellae in a number of sacculi. Absence of the extracellular glycans poly-β-1,6-𝒩-acetylglucosamine and a cellulose-like polymer, produced by the MatAB and CslA proteins respectively, results in a thinner sacculus and absence of lamellae and patches. Extracellular glycans might thus form or lead to the formation of the outer cell wall lamella. Based on these findings we propose a revisited model for the complex cell wall architecture of an apically growing bacterium, in which the network of peptidoglycan together with extracellular polymers is structurally supported by teichoic acids.

1975 ◽  
Vol 21 (1) ◽  
pp. 9-16 ◽  
Author(s):  
D. F. Day ◽  
J. M. Ingram

Alkaline phosphatase (APase) of Pseudomonas aeruginosa exists primarily in the periplasmic region of the cell, i.e., between the cytoplasmic membrane and the outer tripartite layer. The enzyme is also found in the culture filtrate or associated with the outer layer of the cell wall. APase forms a complex with released outer cell wall material, and lipopolysaccharide (LPS) is associated with the complex. Since the enzyme was purified to homogeneity, it became desirable to determine whether complex formation with LPS, or the outer cell wall, affected any properties of the purified phosphatase. The ratio of activities of purified APase with p-nitrophenylphosphate and β-glycerolphosphate as substrates is about 4:1. The ratio of activities with enzyme complexed with LPS is about 1:1. The energy of activation of sucrose or magnesium released enzyme is 9500 cal/mol whereas the values for purified enzyme plus LPS, purified enzyme, purified enzyme plus phosphatidylethanolamine (PE), and purified enzyme plus LPS plus PE range from 3400 to 8700 cal/mol. These changes occur in the physiological temperature range, 27 to 39C, of this organism. Sucrose-released enzyme in the presence of substrate is inactivated at 47C whereas pure enzyme plus substrate is affected at 41C. The addition of LPS, PE, or a combination of both increases the temperature of inactivation from 45 to 51C. The results suggest that certain properties of the purified enzyme differ from those of the enzyme released from whole cells by either sucrose or magnesium resuspension. The addition of cell wall components such as LPS and PE to purified APase restores these properties. The evidence suggests that artificial complex formation changes the environment of the enzyme protein such that the environment now resembles that which exists within the whole cell wall.


2020 ◽  
Author(s):  
Eveline Ultee ◽  
Ariane Briegel ◽  
Dennis Claessen

ABSTRACTThe cell wall is considered an essential component for bacterial survival, providing structural support and protection from environmental insults. Under normal growth conditions, filamentous actinobacteria insert new cell wall material at the hyphal tips regulated by the coordinated activity of cytoskeletal proteins and cell wall biosynthetic enzymes. Despite the importance of the cell wall, some filamentous actinobacteria can produce wall-deficient S-cells upon prolonged exposure to hyperosmotic stress. Here we performed cryo-electron tomography and live cell imaging to further characterize S-cell extrusion in Kitasatospora viridifaciens. We show that exposure to hyperosmotic stress leads to DNA compaction, membrane and S-cell extrusion and thinning of the cell wall at hyphal tips. Additionally, we find that the extrusion of S-cells is abolished in a cytoskeletal mutant strain that lacks the intermediate filament-like protein FilP. Furthermore, micro-aerobic culturing promotes the formation of S-cells in the wild-type, but the limited oxygen still impedes S-cell formation in the ΔfilP mutant. These results demonstrate that S-cell formation is stimulated by oxygen-limiting conditions and dependent on the presence of an intact cytoskeleton.


2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Eveline Ultee ◽  
Lizah T. van der Aart ◽  
Le Zhang ◽  
Dino van Dissel ◽  
Christoph A. Diebolder ◽  
...  

Author(s):  
A. E. Sowers ◽  
E. L. Thurston

Plant stinging emergences exhibit functional similarities in that they all elicit a pain response upon contact. A stinging emergence consists of an elongated stinging cell and a multicellular pedestal (Fig. 1). A recent ultrastructural investigation of these structures has revealed the ontogeny and morphology of the stinging cells differs in representative genera in the four plant families which possess such structures. A unique feature of the stinging cell of Urtica dioica is the presence of a siliceous cell wall in the apical portion of the cell. This rigid region of the cell wall is responsible for producing the needle-like apparatus which penetrates the skin. The stinging cell differentiates the apical bulbous tip early in development and the cell continues growth by intercalary addition of non-silicified wall material until maturity.The uppermost region of the stinging cell wall is entirely composed of silica (Fig. 2, 3) and upon etching with a 3% solution of HF (5 seconds), the silica is partially removed revealing the wall consisting of individualized silica bodies (Fig. 4, 5).


Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Angus E. Rae ◽  
Vivien Rolland ◽  
Rosemary G. White ◽  
Ulrike Mathesius

Abstract Background The formation of infection threads in the symbiotic infection of rhizobacteria in legumes is a unique, fascinating, and poorly understood process. Infection threads are tubes of cell wall material that transport rhizobacteria from root hair cells to developing nodules in host roots. They form in a type of reverse tip-growth from an inversion of the root hair cell wall, but the mechanism driving this growth is unknown, and the composition of the thread wall remains unclear. High resolution, 3-dimensional imaging of infection threads, and cell wall component specific labelling, would greatly aid in our understanding of the nature and development of these structures. To date, such imaging has not been done, with infection threads typically imaged by GFP-tagged rhizobia within them, or histochemically in thin sections. Results We have developed new methods of imaging infection threads using novel and traditional cell wall fluorescent labels, and laser confocal scanning microscopy. We applied a new Periodic Acid Schiff (PAS) stain using rhodamine-123 to the labelling of whole cleared infected roots of Medicago truncatula; which allowed for imaging of infection threads in greater 3D detail than had previously been achieved. By the combination of the above method and a calcofluor-white counter-stain, we also succeeded in labelling infection threads and plant cell walls separately, and have potentially discovered a way in which the infection thread matrix can be visualized. Conclusions Our methods have made the imaging and study of infection threads more effective and informative, and present exciting new opportunities for future research in the area.


2018 ◽  
Vol 15 (8) ◽  
pp. 513
Author(s):  
Ewen Silvester ◽  
Annaleise R. Klein ◽  
Kerry L. Whitworth ◽  
Ljiljana Puskar ◽  
Mark J. Tobin

Environmental contextSphagnum moss is a widespread species in peatlands globally and responsible for a large fraction of carbon storage in these systems. We used synchrotron infrared microspectroscopy to characterise the acid-base properties of Sphagnum moss and the conditions under which calcium uptake can occur (essential for plant tissue integrity). The work allows a chemical model for Sphagnum distribution in the landscape to be proposed. AbstractSphagnum is one the major moss types responsible for the deposition of organic soils in peatland systems. The cell walls of this moss have a high proportion of carboxylated polysaccharides (polygalacturonic acids), which act as ion exchangers and are likely to be important for the structural integrity of the cell walls. We used synchrotron light source infrared microspectroscopy to characterise the acid-base and calcium complexation properties of the cell walls of Sphagnum cristatum stems, using freshly sectioned tissue confined in a flowing liquid cell with both normal water and D2O media. The Fourier transform infrared spectra of acid and base forms are consistent with those expected for protonated and deprotonated aliphatic carboxylic acids (such as uronic acids). Spectral deconvolution shows that the dominant aliphatic carboxylic groups in this material behave as a monoprotic acid (pKa=4.97–6.04). The cell wall material shows a high affinity for calcium, with a binding constant (K) in the range 103.9–104.7 (1:1 complex). The chemical complexation model developed here allows for the prediction of the chemical environment (e.g. pH, ionic content) under which Ca2+ uptake can occur, and provides an improved understanding for the observed distribution of Sphagnum in the landscape.


2010 ◽  
Vol 9 (11) ◽  
pp. 1650-1660 ◽  
Author(s):  
Encarnación Dueñas-Santero ◽  
Ana Belén Martín-Cuadrado ◽  
Thierry Fontaine ◽  
Jean-Paul Latgé ◽  
Francisco del Rey ◽  
...  

ABSTRACT In yeast, enzymes with β-glucanase activity are thought to be necessary in morphogenetic events that require controlled hydrolysis of the cell wall. Comparison of the sequence of the Saccharomyces cerevisiae exo-β(1,3)-glucanase Exg1 with the Schizosaccharomyces pombe genome allowed the identification of three genes that were named exg1 + (locus SPBC1105.05), exg2 + (SPAC12B10.11), and exg3 + (SPBC2D10.05). The three proteins have different localizations: Exg1 is secreted to the periplasmic space, Exg2 is a membrane protein, and Exg3 is a cytoplasmic protein. Characterization of the biochemical activity of the proteins indicated that Exg1 and Exg3 are active only against β(1,6)-glucans while no activity was detected for Exg2. Interestingly, Exg1 cleaves the glucans with an endohydrolytic mode of action. exg1 + showed periodic expression during the cell cycle, with a maximum coinciding with the septation process, and its expression was dependent on the transcription factor Sep1. The Exg1 protein localizes to the septum region in a pattern that was different from that of the endo-β(1,3)-glucanase Eng1. Overexpression of Exg2 resulted in an increase in cell wall material at the poles and in the septum, but the putative catalytic activity of the protein was not required for this effect.


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