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Developing a Novel and Optimized Yeast Model for Human VDAC Research.

Martyna Baranek-Grabińska1, Wojciech Grabiński1, Deborah Musso2

  • 1Department of Bioenergetics, Faculty of Biology, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, 61-614 Poznań, Poland.

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Summary

Yeast VDAC paralogs influence human VDAC complementation. The MET15 gene and hydrogen sulfide levels impact hVDAC3 function, highlighting yeast strain differences in studying voltage-dependent anion channels.

Keywords:
MET15cysteine-depleted variant of hVDAC3human VDAC paralogspor1Δpor2Δ mutantsyeast complementation assayyeast strain genotype

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Area of Science:

  • Mitochondrial Biology and Bioenergetics
  • Molecular and Cellular Physiology
  • Yeast Genetics and Systems Biology

Background:

  • Voltage-dependent anion-selective channels (VDACs) are critical for mitochondrial function and cellular activity integration.
  • Human VDAC paralogs (hVDAC1-3) are studied using heterologous expression in yeast Saccharomyces cerevisiae por1Δ mutants.
  • The role of the second yeast VDAC paralog (yVDAC2) and yeast strain genetic background in these studies is often overlooked.

Purpose of the Study:

  • To investigate the influence of yVDAC2 and yeast cell genotype, including MET15 and its effect on hydrogen sulfide (H2S) levels, on the complementation potential of heterologous hVDAC expression.
  • To analyze the impact of yeast auxotrophic markers on the functional differentiation of hVDAC paralogs, particularly hVDAC3.

Main Methods:

  • Utilized por1Δpor2Δ double mutants and single mutants (por1Δ, por2Δ) derived from two distinct Saccharomyces cerevisiae strains (M3 and BY4741).
  • Performed heterologous expression of human VDAC paralogs (hVDAC1-3) in these yeast mutants.
  • Assessed complementation potential and growth phenotypes, considering the presence of MET15 and its associated H2S production.

Main Results:

  • yVDAC2 potentially contributes to the complementation potential of heterologous hVDAC expression.
  • The complementation of hVDAC paralogs, especially hVDAC3, was significantly influenced by the yeast genetic background, including MET15 expression and H2S levels.
  • Differences in cysteine residue modification, influenced by MET15 and H2S, explained varied hVDAC3 phenotypes between yeast strains M3 and BY4741.

Conclusions:

  • Yeast auxotrophic markers, such as MET15, create distinct backgrounds affecting hVDAC paralog function and modification, particularly for hVDAC3.
  • The presence of yVDAC2 may play a role in modulating the complementation efficiency of hVDACs.
  • The distinct phenotypes observed in M3-Δpor1Δpor2 and BY4741-Δpor1Δpor2 cells underscore their value as models for studying human VDACs, especially redox-sensitive hVDAC3.