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Related Experiment Videos

The yeast DASH complex forms closed rings on microtubules.

J J L Miranda1, Peter De Wulf, Peter K Sorger

  • 1Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA.

Nature Structural & Molecular Biology
|January 11, 2005
PubMed
Summary

The Saccharomyces cerevisiae DASH complex, a key kinetochore protein, forms ring-like structures around microtubules. This assembly is crucial for maintaining spindle integrity during cell division.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • The kinetochore is essential for accurate chromosome segregation during cell division.
  • The DASH complex is a vital microtubule-binding protein within the kinetochore.
  • Understanding the DASH complex's structure and function is key to comprehending cell division.

Purpose of the Study:

  • To characterize the structure and assembly of the Saccharomyces cerevisiae DASH complex.
  • To investigate the interaction of the DASH complex with microtubules.
  • To elucidate the role of DASH complex structures in maintaining spindle integrity.

Main Methods:

  • Coexpression of all ten DASH complex subunits in Escherichia coli.
  • Purification of the recombinant DASH complex.

Related Experiment Videos

  • Electron microscopy (EM) to visualize the structure of DASH alone and bound to microtubules.
  • Analysis of hydrodynamic properties to compare recombinant and native complexes.

    • Main Results:

    • A stable, approximately 210-kDa heterodecameric DASH complex was purified, containing one copy of each subunit.
    • The recombinant DASH complex exhibited identical hydrodynamic properties to the native complex.
    • EM revealed a globular structure for the free DASH complex.
    • DASH complex oligomerized into rings and paired helices that encircled microtubules upon binding.

    Conclusions:

    • The DASH complex can be functionally reconstituted in vitro from its individual subunits.
    • The oligomerization of DASH into collar-like structures around microtubules is a key feature of its interaction.
    • These structures likely play a critical role in ensuring proper microtubule attachment and spindle stability during chromosome segregation.