Genetics, Transcriptional Profiles, and Catalytic Properties of the UDP-Arabinose Mutase Family from Barley

Hsieh, Yves S. Y.; Zhang, Qisen; Yap, Kuok; Shirley, Neil J.; Lahnstein, Jelle; Nelson, Clark J.; Burton, Rachel A.; Millar, A. Harvey; Bulone, Vincent

doi:10.1021/acs.biochem.5b01055

Cited by 15 publications

(32 citation statements)

References 54 publications

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“…This raises the question how ArabAz is incorporated. Fincher and coworkers recently reported on the catalytic properties of an UDP-arabinose mutase (UAM) enzyme in Barley that catalyzes the multistep reversible UDP-Ara p → UDP-Ara f reaction [ 32 ]. They state that a key step in this reaction presumably includes cleavage of the arabinosyl residue from UDP-Ara p , which allows opening of the pyranosyl-ring, formation of the furanose ring, and reconnection of the arabinofuranosyl residue to the UDP molecule.…”

Section: Resultsmentioning

confidence: 99%

Direct imaging of glycans in Arabidopsis roots via click labeling of metabolically incorporated azido-monosaccharides

et al. 2016

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BackgroundCarbohydrates, also called glycans, play a crucial but not fully understood role in plant health and development. The non-template driven formation of glycans makes it impossible to image them in vivo with genetically encoded fluorescent tags and related molecular biology approaches. A solution to this problem is the use of tailor-made glycan analogs that are metabolically incorporated by the plant into its glycans. These metabolically incorporated probes can be visualized, but techniques documented so far use toxic copper-catalyzed labeling. To further expand our knowledge of plant glycobiology by direct imaging of its glycans via this method, there is need for novel click-compatible glycan analogs for plants that can be bioorthogonally labelled via copper-free techniques.ResultsArabidopsis seedlings were incubated with azido-containing monosaccharide analogs of N-acetylglucosamine, N-acetylgalactosamine, l-fucose, and l-arabinofuranose. These azido-monosaccharides were metabolically incorporated in plant cell wall glycans of Arabidopsis seedlings. Control experiments indicated active metabolic incorporation of the azido-monosaccharide analogs into glycans rather than through non-specific absorption of the glycan analogs onto the plant cell wall. Successful copper-free labeling reactions were performed, namely an inverse-electron demand Diels-Alder cycloaddition reaction using an incorporated N-acetylglucosamine analog, and a strain-promoted azide-alkyne click reaction. All evaluated azido-monosaccharide analogs were observed to be non-toxic at the used concentrations under normal growth conditions.ConclusionsOur results for the metabolic incorporation and fluorescent labeling of these azido-monosaccharide analogs expand the possibilities for studying plant glycans by direct imaging. Overall we successfully evaluated five azido-monosaccharide analogs for their ability to be metabolically incorporated in Arabidopsis roots and their imaging after fluorescent labeling. This expands the molecular toolbox for direct glycan imaging in plants, from three to eight glycan analogs, which enables more extensive future studies of spatiotemporal glycan dynamics in a wide variety of plant tissues and species. We also show, for the first time in metabolic labeling and imaging of plant glycans, the potential of two copper-free click chemistry methods that are bio-orthogonal and lead to more uniform labeling. These improved labeling methods can be generalized and extended to already existing and future click chemistry-enabled monosaccharide analogs in Arabidopsis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-016-0907-0) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Direct imaging of glycans in Arabidopsis roots via click labeling of metabolically incorporated azido-monosaccharides

et al. 2016

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“…GT61 family members have characterized functions in transferring arabinose and xylose substitutions onto a 1,4-β-xylan backone (Anders et al, 2012 ). GT75 members are annotated as UDP-Ara mutases (UAM), involved in the conversion of UDP-Arabinopyranose to UDP-Arabinofuranose, which is essential for the generation of the UDP-Ara f substrate for arabinoxylan, arabinogalactan protein, and pectic polysaccharide biosynthesis (Hsieh et al, 2015 ). With the recent finding of arabinoxylan in the papillae produced by barley in response to the attempted penetration of Blumeria graminis f.sp.…”

Section: Publically Available Datasets Highlight Complex Transcriptommentioning

confidence: 99%

The Plant Cell Wall: A Complex and Dynamic Structure As Revealed by the Responses of Genes under Stress Conditions

et al. 2016

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The plant cell wall has a diversity of functions. It provides a structural framework to support plant growth and acts as the first line of defense when the plant encounters pathogens. The cell wall must also retain some flexibility, such that when subjected to developmental, biotic, or abiotic stimuli it can be rapidly remodeled in response. Genes encoding enzymes capable of synthesizing or hydrolyzing components of the plant cell wall show differential expression when subjected to different stresses, suggesting they may facilitate stress tolerance through changes in cell wall composition. In this review we summarize recent genetic and transcriptomic data from the literature supporting a role for specific cell wall-related genes in stress responses, in both dicot and monocot systems. These studies highlight that the molecular signatures of cell wall modification are often complex and dynamic, with multiple genes appearing to respond to a given stimulus. Despite this, comparisons between publically available datasets indicate that in many instances cell wall-related genes respond similarly to different pathogens and abiotic stresses, even across the monocot-dicot boundary. We propose that the emerging picture of cell wall remodeling during stress is one that utilizes a common toolkit of cell wall-related genes, multiple modifications to cell wall structure, and a defined set of stress-responsive transcription factors that regulate them.

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“…However, over-expression of these two genes individually led to a significant increase in resistance. A recent study has shown that both MLOC_64204 and MLOC_6065 have UDP-arabinose mutase (UAM) activity and are able to convert UDP- l -arabinopyranose to UDP- l -arabinofuranose (Hsieh et al, 2016 ). UDP- l -Arabinofuranose is the required form of substrate arabinose for arabinosyltransferases that attach arabinosyl residues to the xylan backbone.…”

Section: Discussionmentioning

confidence: 99%

“…Two members of the GT8 family, termed Glucuronic Acid Substitution of Xylan ( GUX ) 1 and GUX2 , introduce glucuronyl (GlcA) substituents onto the xylan backbone (Mortimer et al, 2010 ; Rennie et al, 2012 ). Members of the GT75 family, which encode UDP-arabinose mutases (UAM) or Reversibly Glycosylated Protein (RGP), are responsible for converting UDP-arabinopyranose to UDP-arabinofuranose, which is the substrate for xylan arabinosyltransferases (Rautengarten et al, 2011 ; Hsieh et al, 2016 ). Some members of the GT61 family have xylan arabinosyltransferase activity that adds arabinosyl residues onto the xylan backbone (Anders et al, 2012 ), while another member of the GT61 family is capable of mediating xylosyl substitution of arabinosyl residues on the xylan backbone (Chiniquy et al, 2012 ).…”

Section: Introductionmentioning

confidence: 99%

Altered Expression of Genes Implicated in Xylan Biosynthesis Affects Penetration Resistance against Powdery Mildew

et al. 2017

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Heteroxylan has recently been identified as an important component of papillae, which are formed during powdery mildew infection of barley leaves. Deposition of heteroxylan near the sites of attempted fungal penetration in the epidermal cell wall is believed to enhance the physical resistance to the fungal penetration peg and hence to improve pre-invasion resistance. Several glycosyltransferase (GT) families are implicated in the assembly of heteroxylan in the plant cell wall, and are likely to work together in a multi-enzyme complex. Members of key GT families reported to be involved in heteroxylan biosynthesis are up-regulated in the epidermal layer of barley leaves during powdery mildew infection. Modulation of their expression leads to altered susceptibility levels, suggesting that these genes are important for penetration resistance. The highest level of resistance was achieved when a GT43 gene was co-expressed with a GT47 candidate gene, both of which have been predicted to be involved in xylan backbone biosynthesis. Altering the expression level of several candidate heteroxylan synthesis genes can significantly alter disease susceptibility. This is predicted to occur through changes in the amount and structure of heteroxylan in barley papillae.

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Genetics, Transcriptional Profiles, and Catalytic Properties of the UDP-Arabinose Mutase Family from Barley

Cited by 15 publications

References 54 publications

Direct imaging of glycans in Arabidopsis roots via click labeling of metabolically incorporated azido-monosaccharides

Direct imaging of glycans in Arabidopsis roots via click labeling of metabolically incorporated azido-monosaccharides

The Plant Cell Wall: A Complex and Dynamic Structure As Revealed by the Responses of Genes under Stress Conditions

Altered Expression of Genes Implicated in Xylan Biosynthesis Affects Penetration Resistance against Powdery Mildew

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