Analysis of a cellulose synthase catalytic subunit from the oomycete pathogen of crops Phytophthora capsici

Abstract Phytophthora capsici Leonian is an important oomycete pathogen of crop vegetables, causing significant economic losses each year. Its cell wall, rich in cellulose, is vital for cellular integrity and for interactions with the host organisms. Predicted cellulose synthase (CesA) proteins are expected to catalyze the polymerization of cellulose, but this has not been biochemically demonstrated in an oomycete. Here, we present the properties of the four newly identified CesA proteins from P. capsici and compare their domain organization with that of CesAs from other lineages. Using a newly constructed glucosyltransferase-deficient variant of Saccharomyces cerevisiae with low residual background activity, we have achieved successful heterologous expression and biochemical characterization of a CesA protein from P. capsici (PcCesA1). Our results demonstrate that the individual PcCesA1 enzyme produces cellobiose as the major reaction product. Co-immunoprecipitation studies and activity assays revealed that several PcCesA proteins interact together to form a complex whose multiproteic nature is most likely required for cellulose microfibril formation. In addition to providing important insights into cellulose synthesis in the oomycetes, our data may assist the longer term identification of cell wall biosynthesis inhibitors to control infection by pathogenic oomycetes.

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Identification of Putative Cell Wall Synthesis Genes in Betula pendula

10.21203/rs.3.rs-33560/v2 ◽

2020 ◽

Author(s):

Song Chen ◽

Xin Lin ◽

Xiyang Zhao ◽

Su Chen

Keyword(s):

Cell Wall ◽

Betula Pendula ◽

Secondary Cell Wall ◽

Plant Cell Wall ◽

Gene Families ◽

Cellulose Synthase ◽

Wall Structure ◽

Cellulose Synthesis ◽

Cell Wall Synthesis ◽

Secondary Cell Wall Synthesis

Abstract BackgroundCellulose is an essential structural component of plant cell wall and is an important resource to produce paper, textiles, bioplastics and other biomaterials. The synthesis of cellulose is among the most important but poorly understood biochemical processes, which is precisely regulated by internal and external cues.ResultsHere, we identified 46 gene models in 7 gene families which encoding cellulose synthase and related enzymes of Betula pendula, and the transcript abundance of these genes in xylem, root, leaf and flower tissues also be determined. Based on these RNA-seq data, we have identified 8 genes that most likely participate in secondary cell wall synthesis, which include 3 cellulose synthase genes and 5 cellulose synthase-like genes. In parallel, a gene co-expression network was also constructed based on transcriptome sequencing.ConclusionsIn this study, we have identified a total of 46 cell wall synthesis genes in B. pendula, which include 8 secondary cell wall synthesis genes. These analyses will help decipher the genetic information of the cell wall synthesis genes, elucidate the molecular mechanism of cellulose synthesis and understand the cell wall structure in B. pendula.

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Updating Insights into the Catalytic Domain Properties of Plant Cellulose synthase (CesA) and Cellulose synthase-like (Csl) Proteins

Molecules ◽

10.3390/molecules26144335 ◽

2021 ◽

Vol 26 (14) ◽

pp. 4335

Author(s):

Gerasimos Daras ◽

Dimitris Templalexis ◽

Fengoula Avgeri ◽

Dikran Tsitsekian ◽

Konstantina Karamanou ◽

...

Keyword(s):

Cell Wall ◽

Plant Cell ◽

Plant Cell Wall ◽

Catalytic Domain ◽

Economic Value ◽

Cellulose Synthase ◽

Protein Isoforms ◽

Research Progress ◽

Cellulose Synthesis ◽

Biotic Stresses

The wall is the last frontier of a plant cell involved in modulating growth, development and defense against biotic stresses. Cellulose and additional polysaccharides of plant cell walls are the most abundant biopolymers on earth, having increased in economic value and thereby attracted significant interest in biotechnology. Cellulose biosynthesis constitutes a highly complicated process relying on the formation of cellulose synthase complexes. Cellulose synthase (CesA) and Cellulose synthase-like (Csl) genes encode enzymes that synthesize cellulose and most hemicellulosic polysaccharides. Arabidopsis and rice are invaluable genetic models and reliable representatives of land plants to comprehend cell wall synthesis. During the past two decades, enormous research progress has been made to understand the mechanisms of cellulose synthesis and construction of the plant cell wall. A plethora of cesa and csl mutants have been characterized, providing functional insights into individual protein isoforms. Recent structural studies have uncovered the mode of CesA assembly and the dynamics of cellulose production. Genetics and structural biology have generated new knowledge and have accelerated the pace of discovery in this field, ultimately opening perspectives towards cellulose synthesis manipulation. This review provides an overview of the major breakthroughs gathering previous and recent genetic and structural advancements, focusing on the function of CesA and Csl catalytic domain in plants.

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Cellulose synthase complexes display distinct dynamic behaviors during xylem transdifferentiation

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1802113115 ◽

2018 ◽

Vol 115 (27) ◽

pp. E6366-E6374 ◽

Cited By ~ 21

Author(s):

Yoichiro Watanabe ◽

Rene Schneider ◽

Sarah Barkwill ◽

Eliana Gonzales-Vigil ◽

Joseph L. Hill ◽

...

Keyword(s):

Cell Wall ◽

Plasma Membrane ◽

Cell Expansion ◽

Secondary Wall ◽

Transition Period ◽

Cellulose Synthase ◽

Wall Thickening ◽

Cellulose Synthesis ◽

Primary Wall ◽

Dynamic Behaviors

In plants, plasma membrane-embedded CELLULOSE SYNTHASE (CESA) enzyme complexes deposit cellulose polymers into the developing cell wall. Cellulose synthesis requires two different sets of CESA complexes that are active during cell expansion and secondary cell wall thickening, respectively. Hence, developing xylem cells, which first undergo cell expansion and subsequently deposit thick secondary walls, need to completely reorganize their CESA complexes from primary wall- to secondary wall-specific CESAs. Using live-cell imaging, we analyzed the principles underlying this remodeling. At the onset of secondary wall synthesis, the primary wall CESAs ceased to be delivered to the plasma membrane and were gradually removed from both the plasma membrane and the Golgi. For a brief transition period, both primary wall- and secondary wall-specific CESAs coexisted in banded domains of the plasma membrane where secondary wall synthesis is concentrated. During this transition, primary and secondary wall CESAs displayed discrete dynamic behaviors and sensitivities to the inhibitor isoxaben. As secondary wall-specific CESAs were delivered and inserted into the plasma membrane, the primary wall CESAs became concentrated in prevacuolar compartments and lytic vacuoles. This adjustment in localization between the two CESAs was accompanied by concurrent decreased primary wall CESA and increased secondary wall CESA protein abundance. Our data reveal distinct and dynamic subcellular trafficking patterns that underpin the remodeling of the cellulose biosynthetic machinery, resulting in the removal and degradation of the primary wall CESA complex with concurrent production and recycling of the secondary wall CESAs.

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A G protein-coupled receptor-like module regulates Cellulose Synthase secretion from the endomembrane system in Arabidopsis

10.1101/2020.10.20.345868 ◽

2020 ◽

Author(s):

Heather E. McFarlane ◽

Daniela Mutwil-Anderwald ◽

Jana Verbančič ◽

Kelsey L. Picard ◽

Timothy E. Gookin ◽

...

Keyword(s):

Cell Wall ◽

Plasma Membrane ◽

Golgi Apparatus ◽

G Protein ◽

Protein Complex ◽

Cellulose Synthase ◽

Cell Wall Integrity ◽

Cellulose Synthesis ◽

Endomembrane System ◽

G Protein Coupled

AbstractCellulose synthesis is essential for plant morphology, water transport and defense, and provides raw material for biomaterials and fuels. Cellulose is produced at the plasma membrane by Cellulose Synthase (CESA) protein complexes (CSCs). CSCs are assembled in the endomembrane system and then trafficked from the Golgi apparatus and trans-Golgi Network (TGN) to the plasma membrane. Since CESA enzymes are only active in the plasma membrane, control of CSC secretion is a critical step in the regulation of cellulose synthesis. However, the regulatory framework for CSC secretion is not clarified. In this study, we identify members of a family of seven transmembrane domain-containing proteins (7TMs) as important for cellulose production during cell wall integrity stress. 7TM proteins are often associated with guanine nucleotide-binding protein (G) protein signalling and mutants in several of the canonical G protein complex components phenocopied the 7tm mutant plants. Unexpectedly, the 7TM proteins localized to the Golgi apparatus/TGN where they interacted with the G protein complex. Here, the 7TMs and G proteins regulated CESA trafficking, but did not affect general protein secretion. Furthermore, during cell wall stress, 7TMs’ localization was biased towards small CESA-containing vesicles, specifically associated with CSC trafficking. Our results thus outline how a G protein-coupled module regulates CESA trafficking and reveal that defects in this process lead to exacerbated responses upon exposure to cell wall integrity stress.

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Cellulose synthase complex organization and cellulose microfibril structure

Philosophical Transactions of The Royal Society A Mathematical Physical and Engineering Sciences ◽

10.1098/rsta.2017.0048 ◽

2017 ◽

Vol 376 (2112) ◽

pp. 20170048 ◽

Cited By ~ 14

Author(s):

Simon Turner ◽

Manoj Kumar

Keyword(s):

Cellulose Microfibril ◽

Current Knowledge ◽

Local Environment ◽

Cellulose Synthase ◽

Cellulose Microfibrils ◽

Cellulose Synthesis ◽

Cellulose Biosynthesis ◽

Link Type ◽

Cellulose Synthase Complex ◽

The Individual

Cellulose consists of linear chains of β-1,4-linked glucose units, which are synthesized by the cellulose synthase complex (CSC). In plants, these chains associate in an ordered manner to form the cellulose microfibrils. Both the CSC and the local environment in which the individual chains coalesce to form the cellulose microfibril determine the structure and the unique physical properties of the microfibril. There are several recent reviews that cover many aspects of cellulose biosynthesis, which include trafficking of the complex to the plasma membrane and the relationship between the movement of the CSC and the underlying cortical microtubules (Bringmann et al. 2012 Trends Plant Sci. 17 , 666–674 ( doi:10.1016/j.tplants.2012.06.003 ); Kumar & Turner 2015 Phytochemistry 112 , 91–99 ( doi:10.1016/j.phytochem.2014.07.009 ); Schneider et al. 2016 Curr. Opin. Plant Biol. 34 , 9–16 ( doi:10.1016/j.pbi.2016.07.007 )). In this review, we will focus on recent advances in cellulose biosynthesis in plants, with an emphasis on our current understanding of the structure of individual catalytic subunits together with the local membrane environment where cellulose synthesis occurs. We will attempt to relate this information to our current knowledge of the structure of the cellulose microfibril and propose a model in which variations in the structure of the CSC have important implications for the structure of the cellulose microfibril produced. This article is part of a discussion meeting issue ‘New horizons for cellulose nanotechnology’.

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Inhibition of cell expansion enhances cortical microtubule stability in the root apex of Arabidopsis thaliana

Journal of Biological Research-Thessaloniki ◽

10.1186/s40709-021-00143-8 ◽

2021 ◽

Vol 28 (1) ◽

Author(s):

Veronica Giourieva ◽

Emmanuel Panteris

Keyword(s):

Arabidopsis Thaliana ◽

Cell Wall ◽

Cell Expansion ◽

Cortical Microtubule ◽

Root Apex ◽

Cellulose Synthase ◽

Cellulose Biosynthesis ◽

Cortical Microtubules ◽

Wild Type ◽

Microtubule Stability

Abstract Background Cortical microtubules regulate cell expansion by determining cellulose microfibril orientation in the root apex of Arabidopsis thaliana. While the regulation of cell wall properties by cortical microtubules is well studied, the data on the influence of cell wall to cortical microtubule organization and stability remain scarce. Studies on cellulose biosynthesis mutants revealed that cortical microtubules depend on Cellulose Synthase A (CESA) function and/or cell expansion. Furthermore, it has been reported that cortical microtubules in cellulose-deficient mutants are hypersensitive to oryzalin. In this work, the persistence of cortical microtubules against anti-microtubule treatment was thoroughly studied in the roots of several cesa mutants, namely thanatos, mre1, any1, prc1-1 and rsw1, and the Cellulose Synthase Interacting 1 protein (csi1) mutant pom2-4. In addition, various treatments with drugs affecting cell expansion were performed on wild-type roots. Whole mount tubulin immunolabeling was applied in the above roots and observations were performed by confocal microscopy. Results Cortical microtubules in all mutants showed statistically significant increased persistence against anti-microtubule drugs, compared to those of the wild-type. Furthermore, to examine if the enhanced stability of cortical microtubules was due to reduced cellulose biosynthesis or to suppression of cell expansion, treatments of wild-type roots with 2,6-dichlorobenzonitrile (DCB) and Congo red were performed. After these treatments, cortical microtubules appeared more resistant to oryzalin, than in the control. Conclusions According to these findings, it may be concluded that inhibition of cell expansion, irrespective of the cause, results in increased microtubule stability in A. thaliana root. In addition, cell expansion does not only rely on cortical microtubule orientation but also plays a regulatory role in microtubule dynamics, as well. Various hypotheses may explain the increased cortical microtubule stability under decreased cell expansion such as the role of cell wall sensors and the presence of less dynamic cortical microtubules.

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METHYL 3-NITRO- AND 3-AMINO-3-DEOXY-β-D-GALACTOPYRANOSIDE AND -MANNOPYRANOSIDE

Canadian Journal of Chemistry ◽

10.1139/v63-219 ◽

1963 ◽

Vol 41 (6) ◽

pp. 1606-1611 ◽

Cited By ~ 26

Author(s):

Hans Helmut Baer ◽

Frank Kienzle

Keyword(s):

Crystalline State ◽

Major Reaction Product ◽

Major Reaction ◽

Amino Derivatives ◽

Facile Preparation ◽

Hydrolysis Of

The steric course of the nitromethane cyclization of L′-methoxy-D-hydroxymethyldiglycolic aldehyde was investigated. Methyl 3-nitro-3-deoxy-β-D-galactopyranoside was shown to arise as a second major reaction product in addition to the previously isolated principal stereoisomer, the gluco derivative. The corresponding manno stereoisomer is formed to a smaller extent. The configurations of the new methyl nitrodeoxyglycosides were established by conversion into the corresponding amino derivatives and hydrolysis of these latter to the known 3-amino-3-deoxy-D-galactose and -D-mannose hydrochlorides. All the products were obtained in a crystalline state. The reaction lends itself to a facile preparation of the nitrogenous galactose derivatives.

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The trafficking and behavior of cellulose synthase and a glimpse of potential cellulose synthesis regulators

Frontiers in Biology ◽

10.1007/s11515-011-1161-3 ◽

2011 ◽

Vol 6 (5) ◽

pp. 377-383 ◽

Cited By ~ 6

Author(s):

Logan Bashline ◽

Juan Du ◽

Ying Gu

Keyword(s):

Cellulose Synthase ◽

Cellulose Synthesis ◽

And Behavior

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Biochemical characterization of a cortexless mutant of a variant of Bacillus cereus

Canadian Journal of Microbiology ◽

10.1139/m76-147 ◽

1976 ◽

Vol 22 (7) ◽

pp. 1007-1012 ◽

Cited By ~ 2

Author(s):

Susanne M. Pearce

Keyword(s):

Cell Wall ◽

Fatty Acid Composition ◽

Fatty Acid ◽

Bacillus Cereus ◽

Biochemical Characterization ◽

Diaminopimelic Acid ◽

Peptidoglycan Synthesis ◽

Biochemical Lesion ◽

Do So

Previous studies on this cortexless mutant of Bacillus cereus var. alesti indicated that the forespore membrane was the site of the biochemical lesion. This hypothesis is supported by the results presented here: fatty acid composition of sporulating cells of the mutant is altered, while in vegetative cells it is comparable to the parent; soluble precursors of peptidoglycan synthesis are accumulated in the mutant, at the time of cortex formation; homogenates of the mutant prepared at the time of cortex formation are unable to incorporate tritiated diaminopimelic acid into peptidoglycan, while homogenates of cells forming germ cell wall do so to an extent comparable to that of the parent; lipid-linked intermediates are formed by the mutant as in the parent. Apparently the mutant is unable either to transfer disaccharide penta-peptide units from the carrier lipid to the growing peptidoglycan acceptor, or to transport lipid-linked intermediates across the forespore membrane.

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