title: Maize plasma membrane aquaporin ZmPIP2;5, but not ZmPIP1;2, facilitates transmembrane diffusion of hydrogen peroxide.
abstract: Plant aquaporins play important roles in transmembrane water transport processes, but some also facilitate the diffusion of other small uncharged solutes ranging from gases to metalloids. Recent evidence suggests that the transmembrane movement of hydrogen peroxide, an intra- and intercellular multifunctional signaling and defense compound, can be regulated by aquaporins. We addressed the question whether maize aquaporins belonging to the plasma membrane intrinsic protein (PIP) subfamily facilitate hydrogen peroxide diffusion using heterologous expression in the yeast Saccharomyces cerevisiae. We showed that ZmPIP proteins belonging to the PIP1 and PIP2 groups were significantly expressed in yeast cells only after codon optimization of their cDNA. In accordance with previous localization studies in oocytes and plants, ZmPIP1;2 was mainly retained in intracellular membranes, while ZmPIP2;5 was localized to the plasma membrane. However, upon co-expression with ZmPIP2;5, ZmPIP1;2 was re-localized to the plasma membrane. Using a non-functional plasma membrane-localized ZmPIP2;5 mutant to deliver ZmPIP1;2 to the plasma membrane, we demonstrated that, in contrast to wild type ZmPIP2;5, ZmPIP1;2 was not permeable to hydrogen peroxide. Our study further highlighted the fact that, when using the yeast system, which is widely employed to study substrates for plant aquaporins and other transporters, although positive transport assay results allow direct conclusions to be drawn regarding solute permeability, negative results require additional control experiments to show that the protein is expressed and localized correctly before concluding on the lack of transport activity.
authors.affiliation: Université catholique de Louvain, Institut des Sciences de la Vie, Croix du Sud, 4-L7.07.14, 1348 Louvain-la-Neuve, Belgium; Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstrasse 3, 06466 Gatersleben, Germany.
authors.forename: Gerd P
authors.initials: GP
authors.lastname: Bienert
authors.forename: Robert B
authors.initials: RB
authors.lastname: Heinen
authors.forename: Marie C
authors.initials: MC
authors.lastname: Berny
authors.forename: François
authors.initials: F
authors.lastname: Chaumont
mesh_terms: D051398:Aquaporin 1 / Q000235:genetics / Q000378:metabolism; D051399:Aquaporin 2 / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D004058:Diffusion; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D006861:Hydrogen Peroxide / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D003313:Zea mays / Q000235:genetics / Q000378:metabolism*
chemical_list: D051399:Aquaporin 2; D026901:Membrane Transport Proteins; D010940:Plant Proteins; D020033:Protein Isoforms; D051398:Aquaporin 1; D006861:Hydrogen Peroxide
keywords: Aquaporin; Hydrogen peroxide; Plant; Plasma membrane intrinsic protein; Plasma membrane localization; Yeast heterologous expression