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Expression of heterologous aquaporins for functional analysis in Saccharomyces cerevisiae

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Abstract

In this study the yeast Saccharomyces cerevisiae, which is a genetically tractable model for analysis of osmoregulation, has been used for analysis of heterologous aquaporins. Aquaporin water channels play important roles in the control of water homeostasis in individual cells and multicellular organisms. We have investigated the effects of functional expression of the mammalian aquaporins AQP1 and AQP5 and the aquaglyceroporins AQP3 and AQP9. Expression of aquaporins caused moderate growth inhibition under hyperosmotic stress, while expression of aquaglyceroporins mediated strong growth inhibition due to glycerol loss. Water transport was monitored in protoplasts, where the kinetics of bursting was influenced by presence of aquaporins but not aquaglyceroporins. We observed glycerol transport through aquaglyceroporins, but not aquaporins, in a yeast strain deficient in glycerol production, whose growth depends on glycerol inflow. In addition, a gene reporter assay allowed to indirectly monitor the effect of AQP9-mediated enhanced glycerol loss on osmoadaptation. Transport activity of certain aqua(glycero)porins was diminished by low pH or CuSO4, suggesting that yeast can potentially be used for screening of putative aquaporin inhibitors. We conclude that yeast is a versatile system for functional studies of aquaporins, and it can be developed to screen for compounds of potential pharmacological use.

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Acknowledgments

We thank members of the group for helpful discussions and for critical reading of the manuscript. This work was supported by grants from the European Commission (contracts BIO4-CT98-0024 and FMRX-CT96-0128 to SH and contract LSHG-CT-2004-504601 to RMB) and the Swedish Research Council (research position and research grant to SH).

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Authors and Affiliations

  1. Department of Cell and Molecular Biology, Göteborg University, Box 462, 40530, Göteborg, Sweden

    Nina Pettersson, Johan Hagström & Stefan Hohmann

  2. School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, B4 7ET, UK

    Roslyn M. Bill

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  1. Nina Pettersson
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  2. Johan Hagström
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  3. Roslyn M. Bill
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  4. Stefan Hohmann
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Correspondence to Stefan Hohmann.

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Communicated by L. Tomaska

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Pettersson, N., Hagström, J., Bill, R.M. et al. Expression of heterologous aquaporins for functional analysis in Saccharomyces cerevisiae . Curr Genet 50, 247–255 (2006). https://doi.org/10.1007/s00294-006-0092-z

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  • DOI: https://doi.org/10.1007/s00294-006-0092-z

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