title: Exploring three PIPs and three TIPs of grapevine for transport of water and atypical substrates through heterologous expression in aqy-null yeast.
abstract: Aquaporins are membrane channels that facilitate the transport of water and other small molecules across the cellular membranes. We examined the role of six aquaporins of Vitis vinifera (cv. Touriga nacional) in the transport of water and atypical substrates (other than water) in an aqy-null strain of Saccharomyces cerevisiae. Their functional characterization for water transport was performed by stopped-flow fluorescence spectroscopy. The evaluation of permeability coefficients (Pf) and activation energies (Ea) revealed that three aquaporins (VvTnPIP2;1, VvTnTIP1;1 and VvTnTIP2;2) are functional for water transport, while the other three (VvTnPIP1;4, VvTnPIP2;3 and VvTnTIP4;1) are non-functional. TIPs (VvTnTIP1;1 and VvTnTIP2;2) exhibited higher water permeability than VvTnPIP2;1. All functional aquaporins were found to be sensitive to HgCl2, since their water conductivity was reduced (24-38%) by the addition of 0.5 mM HgCl2. Expression of Vitis aquaporins caused different sensitive phenotypes to yeast strains when grown under hyperosmotic stress generated by KCl or sorbitol. Our results also indicate that Vitis aquaporins are putative transporters of other small molecules of physiological importance. Their sequence analyses revealed the presence of signature sequences for transport of ammonia, boron, CO2, H2O2 and urea. The phenotypic growth variations of yeast cells showed that heterologous expression of Vitis aquaporins increased susceptibility to externally applied boron and H2O2, suggesting the contribution of Vitis aquaporins in the transport of these species.
authors.affiliation: Centre for Botany Applied to Agriculture (CBAA), Instituto Superior de Agronomia, University of Lisbon, Lisbon, Portugal; Instituto de Investigação do Medicamento (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Farzana
authors.initials: F
authors.lastname: Sabir
authors.affiliation: Centre for Botany Applied to Agriculture (CBAA), Instituto Superior de Agronomia, University of Lisbon, Lisbon, Portugal.
authors.forename: Maria José
authors.initials: MJ
authors.lastname: Leandro
authors.affiliation: Centre for Botany Applied to Agriculture (CBAA), Instituto Superior de Agronomia, University of Lisbon, Lisbon, Portugal; Instituto de Investigação do Medicamento (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Martins
authors.affiliation: Centre for Botany Applied to Agriculture (CBAA), Instituto Superior de Agronomia, University of Lisbon, Lisbon, Portugal.
authors.forename: Maria C
authors.initials: MC
authors.lastname: Loureiro-Dias
authors.affiliation: Instituto de Investigação do Medicamento (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Teresa F
authors.initials: TF
authors.lastname: Moura
authors.affiliation: Instituto de Investigação do Medicamento (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Lisboa, Portugal; Dept. de Bioquímica e Biologia Humana, Faculdade de Farmácia, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Graça
authors.initials: G
authors.lastname: Soveral
authors.affiliation: Centre for Botany Applied to Agriculture (CBAA), Instituto Superior de Agronomia, University of Lisbon, Lisbon, Portugal.
authors.forename: Catarina
authors.initials: C
authors.lastname: Prista
mesh_terms: D020346:Aquaporins / Q000172:deficiency* / Q000235:genetics*; D001692:Biological Transport / Q000187:drug effects; D001895:Boron / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D017353:Gene Deletion; D015870:Gene Expression; D006861:Hydrogen Peroxide / Q000378:metabolism; D008969:Molecular Sequence Data; D009997:Osmotic Pressure / Q000187:drug effects; D011189:Potassium Chloride / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D013012:Sorbitol / Q000494:pharmacology; D027843:Vitis / Q000235:genetics*; D014867:Water / Q000378:metabolism*
chemical_list: D020346:Aquaporins; D014867:Water; D013012:Sorbitol; D011189:Potassium Chloride; D006861:Hydrogen Peroxide; D001895:Boron
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