scholarly journals Biomarker identification of isolated compartments of the cell wall, cytoplasm and vacuole from the internodal cell of characean Nitellopsis obtusa

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e10930
Author(s):  
Brigita Gylytė ◽  
Sigita Jurkonienė ◽  
Reda Cimmperman ◽  
Vaidevutis Šveikauskas ◽  
Levonas Manusadžianas
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Cell Biology ◽  
Model Organism ◽  
Close Relation ◽  
Nitellopsis Obtusa ◽  
Higher Plant ◽  
Large Size ◽  
Enzymatic Markers ◽  
Cell Fractions

Cells of characean algae are attractive for plant cell physiologists because of their large size and their close relation to higher plant cells. The objective of our study was to evaluate the purity of the compartments (cell wall, cytoplasm with plastids, mitochondria, nuclei and endomembrane system, and vacuole) separated mechanically from the internodal cells of Nitellopsis obtusa using enzymatic markers. These included α-mannosidase and malate dehydrogenase, vacuolar and cytoplasmic enzymes, respectively. The biomarkers applied revealed the degree of compartment contamination with the material from unwanted cell parts. The cell wall was contaminated slightly by vacuole and cytoplasm residuals, respectively by 12.3 and 1.96% of corresponding biomarker activities. Relatively high activity of vacuolar marker in the cell wall could be associated with the cell vacuoles in the multicellular structure of the nodes. The biomarkers confirmed highly purified vacuolar (99.5%) and cytoplasmic (86.7%) compartments. Purity estimation of the cell fractions enabled reevaluating nCuO related Cu concentrations in the compartments of charophyte cell. The internalisation of CuO nanoparticles in N. obtusa cell occurred already after 0.5h. In general, the approach seems to be useful for assessing the accumulation and distribution of various xenobiotics and/or metabolites within plant cell. All this justifies N.obtusa internodal cells as a model organism for modern studies in cell biology and nanotoxicology.

Plant Cell Wall Proteomes: Bioinformatics and Cell Biology Tools to Assess the Bona Fide Cell Wall Localization of Proteins

2020 ◽  
pp. 443-462
Author(s):  
David Roujol ◽  
Laurent Hoffmann ◽  
Hélène San Clemente ◽  
Corinne Schmitt-Keichinger ◽  
Christophe Ritzenthaler ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Cell Biology ◽  
Plant Cell Wall ◽  
Bona Fide

scholarly journals Structure and mutagenic analysis of the lipid II flippase MurJ from Escherichia coli

2018 ◽  
Author(s):  
Sanduo Zheng ◽  
Lok-To Sham ◽  
Frederick A. Rubino ◽  
Kelly Brock ◽  
William P. Robins ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Wall ◽  
Plasma Membrane ◽  
Cell Biology ◽  
Model Organism ◽  
Structural Data ◽  
Outer Face ◽  
Antibiotic Development ◽  
Cell Wall Assembly ◽  
Lipid Ii

AbstractThe peptidoglycan cell wall provides an essential protective barrier in almost all bacteria, defining cellular morphology and conferring resistance to osmotic stress and other environmental hazards. The precursor to peptidoglycan, lipid II, is assembled on the inner leaflet of the plasma membrane. However, peptidoglycan polymerization occurs on the outer face of the plasma membrane, and lipid II must be flipped across the membrane by the MurJ protein prior to its use in peptidoglycan synthesis. Due to its central role in cell wall assembly, MurJ is of fundamental importance in microbial cell biology and is a prime target for novel antibiotic development. However, relatively little is known regarding the mechanisms of MurJ function, and structural data are only available for MurJ from the extremophile Thermosipho africanus. Here, we report the crystal structure of substrate-free MurJ from the Gram-negative model organism Escherichia coli, revealing an inward-open conformation. Taking advantage of the genetic tractability of E. coli, we performed high-throughput mutagenesis and next-generation sequencing to assess mutational tolerance at every amino acid in the protein, providing a detailed functional and structural map for the enzyme and identifying sites for inhibitor development. Finally, through the use of sequence co-evolution analysis we identify functionally important interactions in the outward-open state of the protein, supporting a rocker-switch model for lipid II transport.

Root hair cell walls: filling in the frameworkThis review is one of a selection of papers published in the Special Issue on Plant Cell Biology.

2006 ◽  
Vol 84 (4) ◽  
pp. 613-621 ◽  
Author(s):  
M.E. Galway
Keyword(s):  
Cell Wall ◽  
Hair Cell ◽  
Plant Cell ◽  
Root Hair ◽  
Cell Walls ◽  
Cell Biology ◽  
High Speed ◽  
Root Hairs ◽  
Experimental System ◽  
Plant Cell Walls

Rapid progress is being made in determining the composition, synthesis, and mechanical properties of plant cell walls. Although tip-growing root hairs provide an excellent example of high-speed cell wall assembly, they have been relatively neglected by researchers interested in cell walls and those interested in tip growth. This review aims to present the root hair as an experimental system for future cell wall studies by assembling recent discoveries about the walls onto the existing framework based on older information. Most recent data come from arabidopsis ( Arabidopsis thaliana (L.) Heynh) and model legumes. Evidence supporting the turgor-mediated expansion of hair cell walls is considered, along with a survey of three components needed for cell wall expansion without rupture: cellulose (the role of CesA cellulose synthases is also addressed), Csld3, a cellulose synthase-like protein, and Lrx1, a cell wall protein. Further clues about hair cell wall composition have been obtained from gene expression studies and the use of monoclonal antibodies. Finally, there is a review of the experimental evidence that (i) hairs near the hypocotyl differ developmentally and structurally from other hairs and (ii) biosynthesis of wall components in hairs may differ significantly from the epidermal cells that they grew from. All of these recent advances suggest that root hairs could provide valuable data to augment models of plant cell walls based on more conventional cell types.

scholarly journals Mosses: Accessible Systems for Plant Development Studies

2021 ◽  
Author(s):  
Jordi Floriach-Clark ◽  
Han Tang ◽  
Viola Willemsen
Keyword(s):  
Plant Cell ◽  
Cell Biology ◽  
Vascular Plants ◽  
Asymmetric Cell Division ◽  
Apical Cell ◽  
Model Organism ◽  
Land Plant ◽  
Biological Research ◽  
Central System ◽  
Future Potential

Mosses are a cosmopolitan group of land plants, sister to vascular plants, with a high potential for molecular and cell biological research. The species Physcomitrium patens has helped gaining better understanding of the biological processes of the plant cell, and it has become a central system to understand water-to-land plant transition through 2D-to-3D growth transition, regulation of asymmetric cell division, shoot apical cell establishment and maintenance, phyllotaxis and regeneration. P. patens was the first fully sequenced moss in 2008, with the latest annotated release in 2018. It has been shown that many gene functions and networks are conserved in mosses when compared to angiosperms. Importantly, this model organism has a simplified and accessible body structure that facilitates close tracking in time and space with the support of live cell imaging set-ups and multiple reporter lines. This has become possible thanks to its fully established molecular toolkit, with highly efficient PEG-assisted, CRISPR/Cas9 and RNAi transformation and silencing protocols, among others. Here we provide examples on how mosses exhibit advantages over vascular plants to study several processes and their future potential to answer some other outstanding questions in plant cell biology.

scholarly journals Determination of Subunit Composition of Clostridium cellulovorans Cellulosomes That Degrade Plant Cell Walls

2002 ◽  
Vol 68 (4) ◽  
pp. 1610-1615 ◽  
Author(s):  
Koichiro Murashima ◽  
Akihiko Kosugi ◽  
Roy H. Doi
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Scaffolding Protein ◽  
Plant Cell Walls ◽  
Higher Plant ◽  
Clostridium Cellulovorans ◽  
Plant Cell Wall Degradation

ABSTRACT Clostridium cellulovorans produces a cellulase enzyme complex (cellulosome). In this study, we isolated two plant cell wall-degrading cellulosomal fractions from culture supernatant of C. cellulovorans and determined their subunit compositions and enzymatic activities. One of the cellulosomal fractions showed fourfold-higher plant cell wall-degrading activity than the other. Both cellulosomal fractions contained the same nine subunits (the scaffolding protein CbpA, endoglucanases EngE and EngK, cellobiohydrolase ExgS, xylanase XynA, mannanase ManA, and three unknown proteins), although the relative amounts of the subunits differed. Since only cellobiose was released from plant cell walls by the cellulosomal fractions, cellobiohydrolases were considered to be key enzymes for plant cell wall degradation.

scholarly journals Current and future advances in fluorescence-based visualization of plant cell wall components and cell wall biosynthetic machineries

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Brian T DeVree ◽  
Lisa M Steiner ◽  
Sylwia Głazowska ◽  
Felix Ruhnow ◽  
Klaus Herburger ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Cell Walls ◽  
Cell Biology ◽  
Plant Cell Wall ◽  
Imaging Techniques ◽  
Molecular Probes ◽  
Comprehensive Overview ◽  
Biotechnology Research ◽  
High Level

AbstractPlant cell wall-derived biomass serves as a renewable source of energy and materials with increasing importance. The cell walls are biomacromolecular assemblies defined by a fine arrangement of different classes of polysaccharides, proteoglycans, and aromatic polymers and are one of the most complex structures in Nature. One of the most challenging tasks of cell biology and biomass biotechnology research is to image the structure and organization of this complex matrix, as well as to visualize the compartmentalized, multiplayer biosynthetic machineries that build the elaborate cell wall architecture. Better knowledge of the plant cells, cell walls, and whole tissue is essential for bioengineering efforts and for designing efficient strategies of industrial deconstruction of the cell wall-derived biomass and its saccharification. Cell wall-directed molecular probes and analysis by light microscopy, which is capable of imaging with a high level of specificity, little sample processing, and often in real time, are important tools to understand cell wall assemblies. This review provides a comprehensive overview about the possibilities for fluorescence label-based imaging techniques and a variety of probing methods, discussing both well-established and emerging tools. Examples of applications of these tools are provided. We also list and discuss the advantages and limitations of the methods. Specifically, we elaborate on what are the most important considerations when applying a particular technique for plants, the potential for future development, and how the plant cell wall field might be inspired by advances in the biomedical and general cell biology fields.

scholarly journals Structure and mutagenic analysis of the lipid II flippase MurJ fromEscherichia coli

2018 ◽  
Vol 115 (26) ◽  
pp. 6709-6714 ◽  
Author(s):  
Sanduo Zheng ◽  
Lok-To Sham ◽  
Frederick A. Rubino ◽  
Kelly P. Brock ◽  
William P. Robins ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plasma Membrane ◽  
Cell Biology ◽  
Model Organism ◽  
Structural Data ◽  
Outer Face ◽  
Antibiotic Development ◽  
Cell Wall Assembly ◽  
Lipid Ii ◽  
Peptidoglycan Cell Wall

The peptidoglycan cell wall provides an essential protective barrier in almost all bacteria, defining cellular morphology and conferring resistance to osmotic stress and other environmental hazards. The precursor to peptidoglycan, lipid II, is assembled on the inner leaflet of the plasma membrane. However, peptidoglycan polymerization occurs on the outer face of the plasma membrane, and lipid II must be flipped across the membrane by the MurJ protein before its use in peptidoglycan synthesis. Due to its central role in cell wall assembly, MurJ is of fundamental importance in microbial cell biology and is a prime target for novel antibiotic development. However, relatively little is known regarding the mechanisms of MurJ function, and structural data for MurJ are available only from the extremophileThermosipho africanus. Here, we report the crystal structure of substrate-free MurJ from the gram-negative model organismEscherichia coli, revealing an inward-open conformation. Taking advantage of the genetic tractability ofE. coli, we performed high-throughput mutagenesis and next-generation sequencing to assess mutational tolerance at every amino acid in the protein, providing a detailed functional and structural map for the enzyme and identifying sites for inhibitor development. Lastly, through the use of sequence coevolution analysis, we identify functionally important interactions in the outward-open state of the protein, supporting a rocker-switch model for lipid II transport.

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