Analysis of LuPME3, a pectin methylesterase from Linum usitatissimum, revealed a variability in PME proteolytic maturation

Mareck, Alain; Lamour, Romain; Schaumann, Annick; Chan, Philippe; Driouich, Azeddine; Pelloux, Jérôme; Lerouge, Patrice

doi:10.4161/psb.7.1.18632

Cited by 10 publications

(8 citation statements)

References 16 publications

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“…Our transcript abundance data also showed that LuPME5 was expressed in the shoot apex, while LuPME3 was not, which could be correlated with the findings that showed that LuPME5 transcript abundance was higher in the upper parts of the hypocotyl after two days of growth, while LuPME3 was higher in the bottom of the hypocotyl [ 13 ]. Mareck et al [ 14 ] found a very high transcript abundance of LuPME3 in roots, as observed with the promoter fusion in tobacco [ 11 ], in which a GUS construct using LuPME3 promoter was used to detect its expression in stems, roots and leaves. The expression was observed in the vascular tissues of roots, shoots and young leaves.…”

Section: Resultsmentioning

confidence: 99%

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Characterization and transcript profiling of the pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) gene families in flax (Linum usitatissimum)

Pinzón-Latorre

Deyholos

2013

BMC Genomics

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BackgroundPectin methylesterases (PMEs) catalyze the demethylesterification of homogalacturonans in the cell wall; their activity is regulated in part by pectin methylesterase inhibitors (PMEIs). PME activity may result in either rigidification or loosening of the cell wall, depending on the mode of demethylesterification. The activity of PMEs in the middle lamella is expected to affect intrusive elongation of phloem fibers, and their adhesion to adjacent cells. Length and extractability of phloem fibers are qualities important for their industrial uses in textiles and composites. As only three flax PMEs had been previously described, we were motivated to characterize the PME and PMEI gene families of flax.ResultsWe identified 105 putative flax PMEs (LuPMEs) and 95 putative PMEIs (LuPMEIs) within the whole-genome assembly. We found experimental evidence for the transcription of 77/105 LuPMEs and 83/95 LuPMEIs, and surveyed the transcript abundance of these in 12 different tissues and stages of development. Six major monophyletic groups of LuPMEs could be defined based on the inferred relationships of flax genes and their presumed orthologs from other species. We searched the LuPMEs and LuPMEIs for conserved residues previously reported to be important for their tertiary structure and function. In the LuPMEs, the most highly conserved residues were catalytic residues while in the LuPMEIs, cysteines forming disulfude bridges between helices α2 and α3 were most highly conserved. In general, the conservation of critical residues was higher in the genes with evidence of transcript expression than in those for which no expression was detected.ConclusionsThe LuPMEs and LuPMEIs comprise large families with complex patterns of transcript expression and a wide range of physical characteristics. We observed that multiple PMEs and PMEIs are expressed in partially overlapping domains, indicative of several genes acting redundantly during most processes. The potential for functional redundancy was highlighted also by the phylogenetic analyses. We were able to identify a subset of PME and PMEIs that appeared particularly relevant to fiber development, which may provide a basis for the improvement of key traits in industrial feedstocks and a better understanding of the physiological roles of PMEs and PMEIs in general.

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Section: Resultsmentioning

confidence: 99%

“…Three PMEs have been previously characterized in flax: LuPME1, LuPME3, and LuPME5 [ 11 - 14 ]. These are all Type-1 PMEs.…”

Section: Introductionmentioning

confidence: 99%

Characterization and transcript profiling of the pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) gene families in flax (Linum usitatissimum)

Pinzón-Latorre

Deyholos

2013

BMC Genomics

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“…Both PME6 and PME16 are classified as Type I/Group 2 PMEs because they also contain a PRO region with PME inhibitor homology and a processing motif for cleavage by subtilisin-like proteases (Pelloux et al, 2007;Wolf et al, 2009b). While PMEs whose PRO region is not cleaved are predicted to be retained in the Golgi (Wolf et al, 2009b), recent data suggest that certain Type I/Group 2 PMEs can be secreted to the cell wall without processing (Mareck et al, 2012). Therefore, FLY1 could be recycling unprocessed PMEs post-Golgi or upon endocytosis from the PM.…”

Section: Discussionmentioning

confidence: 99%

FLYING SAUCER1 Is a Transmembrane RING E3 Ubiquitin Ligase That Regulates the Degree of Pectin Methylesterification inArabidopsisSeed Mucilage

et al. 2013

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Pectins are complex polysaccharides that form the gel matrix of the primary cell wall and are abundant in the middle lamella that holds plant cells together. Their degree of methylesterification (DM) impacts wall strength and cell adhesion since unesterified pectin regions can cross-link via Ca 2+ ions to form stronger gels. Here, we characterize flying saucer1 (fly1), a novel Arabidopsis thaliana seed coat mutant, which displays primary wall detachment, reduced mucilage extrusion, and increased mucilage adherence. These defects appear to result from a lower DM in mucilage and are enhanced by the addition of Ca 2+ or completely rescued using alkaline Ca 2+ chelators. FLY1 encodes a transmembrane protein with a RING-H2 domain that has in vitro E3 ubiquitin ligase activity. FLY1 is orthologous to TRANSMEMBRANE UBIQUITIN LIGASE1, a Golgi-localized E3 ligase involved in the quality control of membrane proteins in yeast. However, FLY1-yellow fluorescent protein (YFP) fusions are localized in punctae that are predominantly distinct from the Golgi and the trans-Golgi network/early endosome in the seed coat epidermis. Wortmannin treatment, which induces the fusion of late endosomes in plants, resulted in enlarged FLY1-YFP bodies. We propose that FLY1 regulates the DM of pectin in mucilage, potentially by recycling pectin methylesterase enzymes in the endomembrane system of seed coat epidermal cells.

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“…The PME gene encodes a PME precursor containing a variable length N-terminal extension that is essential for protein targeting to the endoplasmic reticulum (Markovic et al, 2002 ). PME maturation requires the removal of the PME leader sequence, including both the transmembrane domain and the spacer sequence (Dorokhov et al, 2006 ; Wolf et al, 2009b ; Mareck et al, 2012 ). The spacer sequence may either function in subcellular targeting by acting as an intramolecular chaperone for the folding of mature enzymes or as an autoinhibitor during transport through the endomembrane system (Pelloux et al, 2007 ; Komarova et al, 2014b ).…”

Section: Introductionmentioning

confidence: 99%

The Intergenic Interplay between Aldose 1-Epimerase-Like Protein and Pectin Methylesterase in Abiotic and Biotic Stress Control

et al. 2017

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The mechanical damage that often precedes the penetration of a leaf by a pathogen promotes the activation of pectin methylesterase (PME); the activation of PME leads to the emission of methanol, resulting in a “priming” effect on intact leaves, which is accompanied by an increased sensitivity to Tobacco mosaic virus (TMV) and resistance to bacteria. In this study, we revealed that mRNA levels of the methanol-inducible gene encoding Nicotiana benthamiana aldose 1-epimerase-like protein (NbAELP) in the leaves of intact plants are very low compared with roots. However, stress and pathogen attack increased the accumulation of the NbAELP mRNA in the leaves. Using transiently transformed plants, we obtained data to support the mechanism underlying AELP/PME-related negative feedback The insertion of the NbAELP promoter sequence (proNbAELP) into the N. benthamiana genome resulted in the co-suppression of the natural NbAELP gene expression, accompanied by a reduction in the NbAELP mRNA content and increased PME synthesis. Knockdown of NbAELP resulted in high activity of PME in the cell wall and a decrease in the leaf glucose level, creating unfavorable conditions for Agrobacterium tumefaciens reproduction in injected leaves. Our results showed that NbAELP is capable of binding the TMV movement protein (MPTMV) in vitro and is likely to affect the cellular nucleocytoplasmic transport, which may explain the sensitivity of NbAELP knockdown plants to TMV. Although NbAELP was primarily detected in the cell wall, the influence of this protein on cellular PME mRNA levels might be associated with reduced transcriptional activity of the PME gene in the nucleus. To confirm this hypothesis, we isolated the N. tabacum PME gene promoter (proNtPME) and showed the inhibition of proNtPME-directed GFP and GUS expression in leaves when co-agroinjected with the NbAELP-encoding plasmid. We hypothesized that plant wounding and/or pathogen attack lead to PME activation and increased methanol emission, followed by increased NbAELP expression, which results in reversion of PME mRNA level and methanol emission to levels found in the intact plant.

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Analysis of LuPME3, a pectin methylesterase from Linum usitatissimum, revealed a variability in PME proteolytic maturation

Cited by 10 publications

References 16 publications

Characterization and transcript profiling of the pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) gene families in flax (Linum usitatissimum)

Characterization and transcript profiling of the pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) gene families in flax (Linum usitatissimum)

FLYING SAUCER1 Is a Transmembrane RING E3 Ubiquitin Ligase That Regulates the Degree of Pectin Methylesterification inArabidopsisSeed Mucilage

The Intergenic Interplay between Aldose 1-Epimerase-Like Protein and Pectin Methylesterase in Abiotic and Biotic Stress Control

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