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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 ( P f  ) and activation energies ( Ea ) revealed that three aquaporins ( Vv TnPIP2;1,  Vv TnTIP1;1 and  Vv TnTIP2;2) are functional for water transport, while the other three ( Vv TnPIP1;4,  Vv TnPIP2;3 and  Vv TnTIP4;1) are non-functional. TIPs ( Vv TnTIP1;1 and  Vv TnTIP2;2) exhibited higher water permeability than  Vv TnPIP2;1. All functional aquaporins were found to be sensitive to HgCl 2 , since their water conductivity was reduced (24–38%) by the addition of 0.5 mM HgCl 2 . 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, CO 2 , H 2 O 2  and urea. The phenotypic growth variations of yeast cells showed that heterologous expression of  Vitis  aquaporins increased susceptibility to externally applied boron and H 2 O 2 , suggesting the contribution of  Vitis  aquaporins in the transport of these species.

Exploring Three PIPs and Three TIPs of Grapevine for Transport of Water and Atypical Substrates through Heterologous Expression in aqy-null Yeast