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Related Experiment Videos

Phosphoregulation and depolymerization-driven movement of the Dam1 complex do not require ring formation.

Daniel R Gestaut1, Beth Graczyk, Jeremy Cooper

  • 1Department of Biochemistry, University of Washington, Seattle, Washington 98195 USA.

Nature Cell Biology
|March 28, 2008
PubMed
Summary
This summary is machine-generated.

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The Dam1 complex, crucial for chromosome segregation, dynamically attaches to microtubules. Phosphorylation by Ipl1 kinase reduces its affinity, promoting detachment and ensuring accurate cell division.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Kinetochores mediate stable microtubule attachments during mitosis.
  • The Dam1 complex is essential for kinetochore-microtubule attachments and is regulated by Ipl1 (Aurora B) kinase.
  • Previous studies suggested Dam1 complex ring formation is key to its function.

Purpose of the Study:

  • To investigate the role of Dam1 complex ring structure in microtubule attachment dynamics.
  • To determine how Ipl1 phosphorylation affects Dam1 complex interaction with microtubules.
  • To elucidate the mechanism of Dam1 complex movement along microtubules.

Main Methods:

  • Utilized two fluorescence-based assays to study Dam1 complex-microtubule interactions.
  • Measured binding affinity (Kd) and detachment frequency following Ipl1 phosphorylation.

Related Experiment Videos

  • Observed Dam1 complex behavior on disassembling microtubule tips.

    • Main Results:

    • Dam1 complex ring formation is not required for dynamic microtubule attachment or Ipl1-dependent affinity modulation.
    • Phosphorylation at Ser 20 by Ipl1 significantly reduces Dam1 complex affinity for microtubules.
    • Individual Dam1 complexes and small oligomers (1-4 units) diffuse along microtubules and are carried by disassembling tips.

    Conclusions:

    • Dynamic, phosphoregulated microtubule attachment can be achieved by small numbers of Dam1 complexes, not requiring ring structures.
    • This mechanism contributes to accurate chromosome segregation by facilitating dynamic kinetochore-microtubule interactions.
    • Ipl1 phosphorylation acts as a key regulator of Dam1 complex function during mitosis.