scholarly journals Formation of wall-less cells in Kitasatospora viridifaciens requires cytoskeletal protein FilP in oxygen-limiting conditions

2020 ◽  
Author(s):  
Eveline Ultee ◽  
Ariane Briegel ◽  
Dennis Claessen
Keyword(s):  
Cell Wall ◽  
Prolonged Exposure ◽  
Electron Tomography ◽  
Cell Formation ◽  
Normal Growth ◽  
Cytoskeletal Proteins ◽  
Hyperosmotic Stress ◽  
Wall Material ◽  
Bacterial Survival ◽  
Cell Extrusion

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.

scholarly journals A MORPHOLOGICAL STUDY OF HALOBACTERIUM HALOBIUM AND ITS LYSIS IN MEDIA OF LOW SALT CONCENTRATION

1967 ◽  
Vol 34 (1) ◽  
pp. 365-393 ◽  
Author(s):  
Walther Stoeckenius ◽  
Robert Rowen
Keyword(s):  
Cell Wall ◽  
Cell Membrane ◽  
Salt Concentration ◽  
Prolonged Exposure ◽  
Structural Characteristics ◽  
Wall Material ◽  
Breakdown Product ◽  
Distilled Water ◽  
Fixation Techniques ◽  
A Cell

The reported absence of a cell wall in halobacteria cannot be confirmed. Improved fixation techniques clearly show a cell wall-like structure on the surface of these cells. A stepwise reduction of the salt concentration causes the release of cell wall material before the cell membrane begins to disintegrate. The cell membrane breaks up into fragments of variable but rather small size, which are clearly different from a 4S component reported by others to be the major breakdown product of the cell membrane. It appears more likely that the 4S component arises from the dissolution of the cell wall. A residue of large membranous sheets remains even after prolonged exposure of halobacteria envelopes to distilled water. The lipids in these sheets do not differ significantly from the lipids in the lysed part of the cell membrane. The sheets, however, contain a purple-colored substance, which is not present in the lysed part. The easily sedimentable residue that remains after lysis of the cells or envelopes in distilled water also contains "intracytoplasmic membranes" with unusual structural characteristics. They can also be identified in sections through intact bacteria or envelope preparations. Their function is at present unknown but seems to be related to the formation of gas vacuoles in these organisms.

scholarly journals Stress-induced formation of cell wall-deficient cells in filamentous actinomycetes

2016 ◽  
Author(s):  
K. Ramijan ◽  
E. Ultee ◽  
J. Willemse ◽  
A.J. Wondergem ◽  
D. Heinrich ◽  
...  
Keyword(s):  
Cell Wall ◽  
Prolonged Exposure ◽  
Gene Clusters ◽  
Adaptation Strategy ◽  
Hyperosmotic Stress ◽  
Cell Type ◽  
Biosynthetic Gene Clusters ◽  
Resistance Determinants ◽  
A Cell ◽  
Almost All

ABSTRACTThe cell wall is a shape-defining structure that envelopes almost all bacteria. One of its main functions is to serve as a protection barrier to environmental stresses. Bacteria can be forced in a cell wall-deficient state under highly specialized conditions, which are invariably aimed at interrupting cell wall synthesis. Therefore, the relevance of such cells has remained obscure. Here we show that many filamentous actinomycetes have a natural ability to generate a new, cell wall-deficient cell type in response to hyperosmotic stress, which we call S-cells. This wall-deficient state is transient, as S-cells are able to switch to the canonical mycelial mode-of-growth. Remarkably, prolonged exposure of S-cells to hyperosmotic stress yielded variants that are able to proliferate indefinitely without their cell wall. This is the first report that demonstrates the formation of wall-deficient cells as a natural adaptation strategy and their potential transition into stable wall-less forms solely caused by prolonged exposure to osmotic stress. Given that actinomycetes are potent antibiotic producers, our work also provides important insights into how biosynthetic gene clusters and resistance determinants may disseminate into the environment.

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 ◽  
...  
Keyword(s):  
Cell Wall ◽  
Electron Tomography ◽  
Streptomyces Coelicolor ◽  
Wall Material ◽  
Outer Cell ◽  
Teichoic Acids ◽  
Structural Support ◽  
Distinct Cell ◽  
Structural Architecture ◽  
Cell Wall Architecture

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.

Silica Incorporation in the Cell Wall of the Singing Cell of Urtica dioica

1975 ◽  
Vol 33 ◽  
pp. 584-585
Author(s):  
A. E. Sowers ◽  
E. L. Thurston
Keyword(s):  
Cell Wall ◽  
Unique Feature ◽  
Urtica Dioica ◽  
Wall Material ◽  
Pain Response ◽  
Apical Portion ◽  
Ultrastructural Investigation ◽  
Plant Families ◽  
Bulbous Tip

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).

scholarly journals New methods for confocal imaging of infection threads in crop and model legumes

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Angus E. Rae ◽  
Vivien Rolland ◽  
Rosemary G. White ◽  
Ulrike Mathesius
Keyword(s):  
Cell Wall ◽  
Root Hair ◽  
Periodic Acid ◽  
Confocal Imaging ◽  
Wall Material ◽  
Future Research ◽  
Thin Sections ◽  
New Methods ◽  
Infection Threads ◽  
Imaging Of Infection

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.

Synchrotron infrared microspectroscopy reveals the response of Sphagnum cell wall material to its aqueous chemical environment

2018 ◽  
Vol 15 (8) ◽  
pp. 513
Author(s):  
Ewen Silvester ◽  
Annaleise R. Klein ◽  
Kerry L. Whitworth ◽  
Ljiljana Puskar ◽  
Mark J. Tobin
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Chemical Environment ◽  
Wall Material ◽  
Organic Soils ◽  
Cell Wall Material ◽  
Acid Base ◽  
Widespread Species ◽  
Flowing Liquid ◽  
Infrared Microspectroscopy

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.

scholarly journals Characterization of Glycoside Hydrolase Family 5 Proteins in Schizosaccharomyces pombe

2010 ◽  
Vol 9 (11) ◽  
pp. 1650-1660 ◽  
Author(s):  
Encarnación Dueñas-Santero ◽  
Ana Belén Martín-Cuadrado ◽  
Thierry Fontaine ◽  
Jean-Paul Latgé ◽  
Francisco del Rey ◽  
...  
Keyword(s):  
Cell Wall ◽  
Schizosaccharomyces Pombe ◽  
Protein Characterization ◽  
Wall Material ◽  
Glycoside Hydrolase Family ◽  
Content Type ◽  
Hydrolysis Of ◽  
Controlled Hydrolysis ◽  
Hydrolase Family

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.

scholarly journals Putative PmrA and PmcA are important for normal growth, morphogenesis and cell wall integrity, but not for viability in Aspergillus nidulans

Microbiology ◽  
2014 ◽  
Vol 160 (11) ◽  
pp. 2387-2395 ◽  
Author(s):  
Hechun Jiang ◽  
Feifei Liu ◽  
Shizhu Zhang ◽  
Ling Lu
Keyword(s):  
Cell Wall ◽  
Plasma Membrane ◽  
Aspergillus Nidulans ◽  
Normal Growth ◽  
Cell Wall Integrity ◽  
Plasma Membrane Atpase ◽  
Growth Defects ◽  
Double Deletion ◽  
Intracellular Ca ◽  
P Type

P-type Ca2+-transporting ATPases are Ca2+ pumps, extruding cytosolic Ca2+ to the extracellular environment or the intracellular Ca2+ store lumens. In budding yeast, Pmr1 (plasma membrane ATPase related), and Pmc1 (plasma membrane calcium-ATPase) cannot be deleted simultaneously for it to survive in standard medium. Here, we deleted two putative Ca2+ pumps, designated AnPmrA and AnPmcA, from Aspergillus nidulans, and obtained the mutants ΔanpmrA and ΔanpmcA, respectively. Then, using ΔanpmrA as the starting strain, the promoter of its anpmcA was replaced with the alcA promoter to secure the mutant ΔanpmrAalcApmcA or its anpmcA was deleted completely to produce the mutant ΔanpmrAΔpmcA. Different from the case in Saccharomyces cerevisiae, double deletion of anpmrA and anpmcA was not lethal in A. nidulans. In addition, deletion of anpmrA and/or anpmcA had produced growth defects, although overexpression of AnPmc1 in ΔanpmrAalcApmcA could not restore the growth defects that resulted from the loss of AnPmrA. Moreover, we found AnPmrA was indispensable for maintenance of normal morphogenesis, especially in low-Ca2+/Mn2+ environments. Thus, our findings suggest AnPmrA and AnPmcA might play important roles in growth, morphogenesis and cell wall integrity in A. nidulans in a different way from that in yeasts.

scholarly journals Reconstruction ofmreBExpression in Staphylococcus aureus via a Collection of New Integrative Plasmids

2014 ◽  
Vol 80 (13) ◽  
pp. 3868-3878 ◽  
Author(s):  
Ana Yepes ◽  
Gudrun Koch ◽  
Andrea Waldvogel ◽  
Juan-Carlos Garcia-Betancur ◽  
Daniel Lopez
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Cell Biology ◽  
Genetic Manipulation ◽  
Protein Localization ◽  
Antibiotic Resistance Genes ◽  
Wall Material ◽  
Content Type ◽  
Genetic Manipulations ◽  
Discrete Structures

ABSTRACTProtein localization has been traditionally explored in unicellular organisms, whose ease of genetic manipulation facilitates molecular characterization. The two rod-shaped bacterial modelsEscherichia coliandBacillus subtilishave been prominently used for this purpose and have displaced other bacteria whose challenges for genetic manipulation have complicated any study of cell biology. Among these bacteria is the spherical pathogenic bacteriumStaphylococcus aureus. In this report, we present a new molecular toolbox that facilitates gene deletion in staphylococci in a 1-step recombination process and additional vectors that facilitate the insertion of diverse reporter fusions into newly identified neutral loci of theS. aureuschromosome. Insertion of the reporters does not add any antibiotic resistance genes to the chromosomes of the resultant strains, thereby making them amenable for further genetic manipulations. We used this toolbox to reconstitute the expression ofmreBinS. aureus, a gene that encodes an actin-like cytoskeletal protein which is absent in coccal cells and is presumably lost during the course of speciation. We observed that inS. aureus, MreB is organized in discrete structures in association with the membrane, leading to an unusual redistribution of the cell wall material. The production of MreB also caused cell enlargement, but it did not revert staphylococcal shape. We present interactions of MreB with key staphylococcal cell wall-related proteins. This work facilitates the useS. aureusas a model system in exploring diverse aspects of cellular microbiology.

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