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Human gasdermin D and MLKL disrupt mitochondria, endocytic traffic and TORC1 signalling in budding yeast

Marta Valenti1, María Molina1, Víctor J Cid1

  • 1Departamento de Microbiología y Parasitología, Facultad de Farmacia, and Instituto Ramón y Cajal de Investigaciones Sanitarias (IRYCIS), Universidad Complutense de Madrid, Madrid 28040, Spain.

Open Biology
|May 23, 2023
PubMed

Insights

Gasdermin D (GSDMD) and mixed lineage kinase domain-like protein (MLKL) disrupt yeast cell functions, including growth and organelle integrity. This study establishes a humanized yeast model for studying these cell death proteins and aiding drug discovery.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Gasdermin D (GSDMD) and mixed lineage kinase domain-like protein (MLKL) are key effectors in pyroptosis and necroptosis, respectively.
  • These proteins induce regulated cell death by disrupting plasma membrane integrity.
  • Budding yeast (Saccharomyces cerevisiae) serves as a model eukaryote for studying human disease-associated proteins via heterologous expression.

Purpose of the Study:

  • To characterize the cellular effects of GSDMD and its N-terminal domain (GSDMD(NT)) in yeast.
  • To compare the effects of GSDMD(NT) with those of MLKL in the same model system.
  • To establish a humanized yeast platform for studying GSDMD and MLKL functions and for drug discovery.

Main Methods:

  • Heterologous expression of GSDMD, GSDMD(NT), and MLKL in Saccharomyces cerevisiae.
  • Analysis of cellular effects including growth inhibition, aggregate formation, mitochondrial morphology, and cell cycle progression.
  • Investigation of TORC1 signaling pathway and vesicular trafficking (endosomal and autophagic).

Main Results:

  • Both GSDMD(NT) and MLKL inhibited yeast growth and formed cytoplasmic aggregates.
  • Mitochondrial fragmentation was observed, with loss-of-function GSDMD(NT) mutants showing organelle affinity.
  • Irreversible cell cycle arrest occurred via TORC1 inhibition, and both proteins disrupted vesicular traffic.

Conclusions:

  • GSDMD and MLKL induce significant cellular perturbations in yeast, mirroring aspects of their function in mammalian cells.
  • The study validates a humanized yeast model for investigating GSDMD and MLKL.
  • This platform is suitable for structure-function analyses and the screening of potential therapeutic compounds.
Keywords:
MLKLSaccharomyces cerevisiaeTORC1endosomal trafficgasdermin Dhumanized yeast

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