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Regulation of the inositol transporter Itr1p by hydrogen peroxide in Saccharomyces cerevisiae

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Abstract

Myo-inositol is a precursor of several membrane phospholipids and sphingolipids and plays a key role in gene regulation in Saccharomyces cerevisiae (S. cerevisiae). Here, we tested whether H2O2 was affecting the levels of the inositol transporters and thus inositol uptake. In S. cerevisiae cells adapted to H2O2 Itr1–GFPp accumulated in the plasma membrane until 20 min, concomitantly with an inhibition of its internalization. Exposure to H2O2 did not alter Itr2–GFPp cellular levels and induced only an 8% decrease at 10 min in the plasma membrane. Therefore, decreased inositol intracellular levels are not caused by decreased levels of inositol transporters in the plasma membrane. However, results show that H2O2 adaptation affects Itr1p turnover and, consequently, H2O2-adapted yeast cells display an inositol transporter phenotype comparable to cells grown in the absence of inositol in growth medium, i.e. accumulation in the plasma membrane and decreased degradation.

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Abbreviations

ANOVA:

One-way analysis of variance

Art5p:

Arrestin-related trafficking adaptors

GFP–Opi1p:

Fusion between green fluorescence protein and Opi1 protein

H2O2 :

Hydrogen peroxide

INO1 :

Inositol-1-phosphate synthase gene

INO2 :

Ino2p gene

ITR1 :

Myo-inositol transporter 1 gene

Itr1–GFPp:

Fusion between Itr1 protein and green fluorescence protein

ITR2 :

Myo-inositol transporter 2 gene

Itr2–GFPp:

Fusion between Itr2 protein and green fluorescence protein

UASINO :

Inositol upstream activating sequence

Rsp5p:

E3 ubiquitin ligase that conjugates ubiquitin to proteins to target them for degradation

SC:

Synthetic complete medium

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Acknowledgements

We thank Dr. André Bastos for help in the endocytosis assays, Dr. Filipe Vilas-Boas for helpful discussions and critical reading of the manuscript and Dr. Helena Soares for help in the preparation of the figures. This work was supported by Fundação para a Ciência e a Tecnologia (FCT), Grant PEst-OE/QUI/UI0612/2013.

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Authors and Affiliations

  1. Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016, Lisbon, Portugal

    Tomás Santos, H. Susana Marinho & Luisa Cyrne

  2. Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016, Lisbon, Portugal

    H. Susana Marinho & Luisa Cyrne

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  1. Tomás Santos
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  2. H. Susana Marinho
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  3. Luisa Cyrne
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Correspondence to Luisa Cyrne.

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Communicated by Olaf Kniemeyer.

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Santos, T., Marinho, H.S. & Cyrne, L. Regulation of the inositol transporter Itr1p by hydrogen peroxide in Saccharomyces cerevisiae. Arch Microbiol 201, 123–134 (2019). https://doi.org/10.1007/s00203-018-1584-1

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  • DOI: https://doi.org/10.1007/s00203-018-1584-1

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