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The Role of LAM Genes in the Pheromone-Induced Cell Death of S. cerevisiae Yeast

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Abstract

Lam1–4 proteins perform non-vesicular transport of sterols from the plasma membrane to the endoplasmic reticulum. Disruption of their function leads to an increase in the content of sterols in the plasma membrane. In mammals, homologs of Lam proteins are responsible for the internalization of plasma cholesterol. The biological role of Lam proteins in yeast remains unclear, since the strains lacking individual LAM genes do not display any pronounced phenotype. Deletion of LAM1 (YSP1) gene inhibits the regulated death of Saccharomyces cerevisiae yeast cells induced by the mating pheromone. Here, we investigated whether LAM2 also plays a role in the cell death induced by the excess of mating pheromone and assessed genetic interactions between LAM2 and genes responsible for ergosterol biosynthesis. We have shown that LAM2 deletion partially prevents pheromone-induced death of yeast cells of the laboratory strain W303, while deletions of three other LAM genes — LAM1, LAM3, and LAM4 — does not provide any additional rescuing effect. The UPC2-1 mutation in the transcription factor UPC2 gene, which leads to the excessive accumulation of sterols in the cell, promotes cell survival in the presence of the pheromone and shows additivity with the LAM2 deletion. On the contrary, LAM2 deletion stimulates pheromone-induced cell death in the laboratory strain BY4741. We have found that the deletion of ergosterol biosynthesis genes ERG2 and ERG6 reduces the effect of LAM2 deletion. Deletion of LAM2 in the Aerg4 strain lacking the gene of the last step of ergosterol biosynthesis, significantly increased the proportion of dead cells and decreased the growth rate of the yeast suspension culture even in the absence of the pheromone. We suggest that the absence of the effect of LAM2 deletion in the Aerg6 and Aerg2 strains indicates the inability of Lam2p to transport some ergosterol biosynthesis intermediates, such as lanosterol. Taken together, our data suggest that the role of Lam proteins in the regulated death of yeast cells caused by the mating pheromone is due to their effect on the plasma membrane sterol composition.

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Abbreviations

CFU:

colony-forming unit

ER:

endoplasmic reticulum

PI:

propidium iodide

PM:

plasma membrane

RCD:

regulated cell death

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Acknowledgements

We thank Vasilina Efimova for participating in the generation of the double AergAlaml mutants.

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Authors and Affiliations

  1. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    S. S. Sokolov, K. V. Galkina, D. A. Knorre & F. F. Severin

  2. Faculty of Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    E. A. Litvinova

  3. Sechenov First Moscow State Medical University, 119991, Moscow, Russia

    D. A. Knorre

Authors
  1. S. S. Sokolov
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  2. K. V. Galkina
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  3. E. A. Litvinova
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  4. D. A. Knorre
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  5. F. F. Severin
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Corresponding author

Correspondence to S. S. Sokolov.

Additional information

Published in Russian in Biokhimiya, 2020, Vol. 85, No. 3, pp. 348–359.

Funding

This work was supported by the Russian Science Foundation (project 18-14-00151) (Figs. 1, 2, and 4–6) and Russian Foundation for Basic Research (project 18-04-01183) (Fig. 3).

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The authors declare no conflict of interest in financial or any other sphere.

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This article does not contain description of studies involving humans or animals as research subjects performed by any of the authors.

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Sokolov, S.S., Galkina, K.V., Litvinova, E.A. et al. The Role of LAM Genes in the Pheromone-Induced Cell Death of S. cerevisiae Yeast. Biochemistry Moscow 85, 300–309 (2020). https://doi.org/10.1134/S0006297920030050

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