The grape aquaporin VvSIP1 transports water across the ER membrane

Henrique Noronha, Alice Agasse, Ana Paula Martins, Marie C. Berny, Dulceneia Gomes, Olfa Zarrouk, Pierre Thiebaud, Serge Delrot, Graça Soveral, François Chaumont, Hernâni Gerós

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)

Abstract

Water diffusion through biological membranes is facilitated by aquaporins, members of the widespread major intrinsic proteins (MIPs). In the present study, the localization, expression, and functional characterization of a small basic intrinsic protein (SIP) from the grapevine were assessed. VvSIP1 was expressed in leaves and berries from field-grown vines, and in leaves and stems from in vitro plantlets, but not in roots. When expressed in tobacco mesophyll cells and in Saccharomyces cerevisiae, fluorescent-tagged VvSIP1 was localized at the endoplasmic reticulum (ER). Stopped-flow spectroscopy showed that VvSIP1-enriched ER membrane vesicles from yeast exhibited higher water permeability and lower activation energy for water transport than control vesicles, indicating the involvement of protein-mediated water diffusion. This aquaporin was able to transport water but not glycerol, urea, sorbitol, glucose, or inositol. VvSIP1 expression in Xenopus oocytes failed to increase the water permeability of the plasma membrane. VvSIP1-His-tag was solubilized and purified to homogeneity from yeast ER membranes and the reconstitution of the purified protein in phosphatidylethanolamine liposomes confirmed its water channel activity. To provide further insights into gene function, the expression of VvSIP1 in mature grapes was studied when vines were cultivated in different field conditions, but its transcript levels did not increase significantly in water-stressed plants and western-exposed berries. However, the expression of the aquaporin genes VvSIP1, VvPIP2;2, and VvTIP1;1 was up-regulated by heat in cultured cells.

Original languageEnglish
Pages (from-to)981-993
Number of pages13
JournalJournal Of Experimental Botany
Volume65
Issue number4
DOIs
Publication statusPublished - Mar 2014

Keywords

  • Aquaporin
  • protein purification
  • proteoliposomes
  • Vitis vinifera
  • VvSIP1
  • water transport.

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