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Characterization of MLKL-mediated Plasma Membrane Rupture in Necroptosis
08:55

Characterization of MLKL-mediated Plasma Membrane Rupture in Necroptosis

Published on: August 7, 2018

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Hsp90 modulates the stability of MLKL and is required for TNF-induced necroptosis

X M Zhao1, Z Chen1, J B Zhao1

  • 1State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Signaling Network, School of Life Sciences, Xiamen University, Xiamen, Fujian, China.

Cell Death & Disease
|February 12, 2016
PubMed

Insights

The molecular chaperone Hsp90 is essential for necroptosis execution by stabilizing key proteins like MLKL and RIP3. Inhibition of Hsp90 disrupts necrosome formation and prevents TNF-induced cell death.

Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Immunology

Background:

  • Mixed lineage kinase domain-like protein (MLKL) is critical for tumor necrosis factor (TNF)-induced necroptosis.
  • Mechanisms regulating MLKL activity and necroptosis execution are not fully understood.

Purpose of the Study:

  • To identify novel regulators of MLKL and necroptosis.
  • To elucidate the role of molecular chaperones in necroptosis.

Main Methods:

  • Co-immunoprecipitation to identify MLKL-interacting proteins.
  • Western blotting to assess protein stability and degradation.
  • Inhibitor studies using 17AAG to disrupt Hsp90 function.
  • Analysis of necrosome assembly and MLKL phosphorylation.
  • Cell viability assays to measure TNF-induced necroptosis.

Main Results:

  • Hsp90 directly interacts with MLKL and is required for its stability.
  • Hsp90 also stabilizes the upstream kinase RIP3.
  • Hsp90 inhibition leads to proteasomal degradation of MLKL and RIP3.
  • Hsp90 is essential for TNF-stimulated necrosome assembly and MLKL phosphorylation.
  • Disruption of Hsp90 function inhibits TNF-induced necroptosis.

Conclusions:

  • Hsp90 is a novel and essential regulator of necroptosis.
  • Hsp90 stabilizes key necroptosis components MLKL and RIP3, promoting their function.
  • Targeting Hsp90 could be a strategy to modulate necroptosis-related pathways.

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