title: Identification and functional characterization of the distinct plant pectin esterases PAE8 and PAE9 and their deletion mutants.
abstract: PAE8 and PAE9 have pectin acetylesterase activity and together remove one-third of the cell wall acetate associated with pectin formation in Arabidopsis leaves. In pae8 and pae9 mutants, substantial amounts of acetate accumulate in cell walls. In addition, the inflorescence stem height is decreased. Pectic polysaccharides constitute a significant part of the primary cell walls in dicotyledonous angiosperms. This diverse group of polysaccharides has been implicated in several physiological processes including cell-to-cell adhesion and pathogenesis. Several pectic polysaccharides contain acetyl-moieties directly affecting their physical properties such as gelling capacity, an important trait for the food industry. In order to gain further insight into the biological role of pectin acetylation, a reverse genetics approach was used to investigate the function of genes that are members of the Pectin AcetylEsterase gene family (PAE) in Arabidopsis. Mutations in two members of the PAE family (PAE8 and PAE9) lead to cell walls with an approximately 20 % increase in acetate content. High-molecular-weight fractions enriched in pectic rhamnogalacturonan I (RGI) extracted from the mutants had increased acetate content. In addition, the pae8 mutant displayed increased acetate content also in low-molecular-weight pectic fractions. The pae8/pae9-2 double mutant exhibited an additive effect by increasing wall acetate content by up to 37 %, suggesting that the two genes are not redundant and act on acetyl-substituents of different pectic domains. The pae8 and pae8/pae9-2 mutants exhibit reduced inflorescence growth underscoring the role of pectic acetylation in plant development. When heterologously expressed and purified, both gene products were shown to release acetate from the corresponding mutant pectic fractions in vitro. PAEs play a significant role in modulating the acetylation state of pectic polymers in the wall, highlighting the importance of apoplastic metabolism for the plant cell and plant growth.
authors.affiliation: Department of Plant and Microbial Biology, Energy Biosciences Institute, University of California, Energy Biosciences Building 212C, 2151 Berkeley Way, Berkeley, CA, 94720-5230, USA.
authors.forename: Amancio
authors.initials: A
authors.lastname: de Souza
authors.forename: Philip A
authors.initials: PA
authors.lastname: Hull
authors.forename: Sascha
authors.initials: S
authors.lastname: Gille
authors.forename: Markus
authors.initials: M
authors.lastname: Pauly
mesh_terms: D000085:Acetates / Q000378:metabolism; D000107:Acetylation; D000115:Acetylesterase / Q000235:genetics* / Q000378:metabolism; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D002265:Carboxylic Ester Hydrolases / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D002473:Cell Wall / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D006603:Hexuronic Acids / Q000378:metabolism; D007527:Isoenzymes / Q000145:classification / Q000235:genetics / Q000378:metabolism; D009005:Monosaccharides / Q000378:metabolism; D009154:Mutation; D010368:Pectins / Q000378:metabolism; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012210:Rhamnose / Q000378:metabolism
chemical_list: D000085:Acetates; D029681:Arabidopsis Proteins; D006603:Hexuronic Acids; D007527:Isoenzymes; D009005:Monosaccharides; C007819:galacturonic acid; D010368:Pectins; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase; D000115:Acetylesterase; C000603269:PAE8 protein, Arabidopsis; C000603268:PAE9 protein, Arabidopsis; D012210:Rhamnose