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Reconstitution of Basic Mitotic Spindles in Spherical Emulsion Droplets
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The Clathrin-dependent Spindle Proteome.

Sushma R Rao1, Neftali Flores-Rodriguez1, Scott L Page1

  • 1From the ‡Children's Medical Research Institute, The University of Sydney, 214 Hawkesbury Road, Westmead, NSW, Australia.

Molecular & Cellular Proteomics : MCP
|May 14, 2016
PubMed
Summary
This summary is machine-generated.

Clathrin stabilizes the mitotic spindle and aids chromosome alignment. This study reveals clathrin

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Proteomics

Background:

  • The mitotic spindle is crucial for accurate chromosome segregation during cell division.
  • Clathrin, an endocytic protein, has a known role in stabilizing the mitotic spindle.
  • The specific molecular pathways clathrin utilizes for spindle stabilization remain largely unknown.

Purpose of the Study:

  • To investigate the signaling pathways involved in clathrin-mediated mitotic spindle stability.
  • To identify proteins and phosphoproteins at the mitotic spindle affected by clathrin depletion.
  • To uncover novel roles of clathrin during mitosis.

Main Methods:

  • Quantitative mass spectrometry (MS/MS) was used to analyze the mitotic spindle proteome and phosphoproteome.
  • Cells were depleted of clathrin to assess its impact on spindle-associated proteins and phosphosites.
  • Proteomic data were analyzed using functional pathway analysis and validated by immunofluorescence microscopy.

Main Results:

  • A comprehensive spindle proteome (3046 proteins) and phosphoproteome (5157 phosphosites) were identified.
  • Clathrin depletion led to the dysregulation of 121 proteins and altered signaling at 47 phosphosites.
  • Functional analysis confirmed clathrin's role in spindle stabilization and chromosome alignment, and revealed a novel role in bipolar spindle formation.

Conclusions:

  • Clathrin plays a critical role in mitotic spindle stabilization and chromosome alignment.
  • The study identified key proteins and signaling pathways regulated by clathrin at the spindle.
  • A novel function of clathrin in bipolar spindle formation during mitosis was discovered.