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Methods to Classify Cytoplasmic Foci as Mammalian Stress Granules
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Methods to Classify Cytoplasmic Foci as Mammalian Stress Granules

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Epithelial Cell Transforming 2 and Aurora Kinase B Modulate Formation of Stress Granule-Containing Transcripts from

Adrienne Weeks1, Sameer Agnihotri2, Jennifer Lymer2

  • 1Division of Neurosurgery, Arthur and Sonia Labatt Brain Tumour Research Centre, Hospital for Sick Children, Toronto, Ontario, Canada; Division of Neurosurgery, Department of Surgery, Dalhousie University, Halifax, Nova Scotia, Canada.

The American Journal of Pathology
|April 24, 2016
PubMed

Insights

Stress granules, containing specific mRNAs, form in astrocytoma cells and are regulated by Aurora kinase B (AurkB). Inhibiting AurkB reduces stress granule formation, suggesting a novel epigenetic regulation target for glioma therapies.

Area of Science:

  • Cell Biology
  • Molecular Oncology
  • Epigenetics

Background:

  • Stress granules are dynamic RNA-protein complexes crucial for cellular stress response and survival.
  • Rho GTPase RhoA and cytokinetic proteins like epithelial cell transforming 2 and Aurora kinase B (AurkB) are implicated in stress granule formation.
  • Astrocytoma cells exhibit unique stress granule compositions relevant to glioma pathogenesis.

Purpose of the Study:

  • To investigate the role of specific cytokinetic proteins, particularly AurkB, in stress granule formation within human astrocytoma cells.
  • To identify the mRNA content of astrocytoma stress granules and their potential involvement in glioma signaling pathways.
  • To explore RNA stress granules as a novel epigenetic regulatory mechanism and potential therapeutic target in astrocytoma.

Main Methods:

  • Localization studies of epithelial cell transforming 2 and AurkB in stress granules.
  • Assessment of AurkB and histone-3 phosphorylation during arsenite-induced stress.
  • High-throughput cellomics assays to quantify stress granule formation after AurkB inhibition (chemical and siRNA).
  • RNA immunoprecipitation followed by mRNA analysis to determine stress granule contents.

Main Results:

  • Epithelial cell transforming 2 and AurkB localize to stress granules in astrocytoma cells.
  • AurkB and histone-3 phosphorylation occur during cellular stress.
  • Inhibition of AurkB leads to a significant reduction in stress granule number and size.
  • Astrocytoma stress granules contain unique mRNAs involved in cellular migration, metabolism, translation, transcriptional regulation, glioma signaling, and the mTOR pathway.

Conclusions:

  • RNA stress granules represent a novel form of epigenetic regulation in astrocytoma.
  • Aurora kinase B plays a critical role in the formation of these stress granules.
  • Targeting RNA stress granules with chemical inhibitors may enhance astrocytoma susceptibility to conventional therapies like radiation and chemotherapy.

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