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Related Experiment Videos

Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function.

D M Brady1, K G Hardwick

  • 1Wellcome Trust Centre for Cell Biology, Institute of Cell and Molecular Biology, University of Edinburgh, UK.

Current Biology : CB
|June 6, 2000
PubMed
Summary

The spindle checkpoint ensures proper cell division by delaying the cell cycle when chromosomes are misaligned. This study reveals a crucial protein complex involving Mad1p, Bub1p, and Bub3p that is essential for this checkpoint

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The spindle checkpoint of budding yeast depends on a tight complex between the Mad1 and Mad2 proteins.

Molecular biology of the cell·1999

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • The spindle checkpoint prevents errors during cell division by monitoring kinetochore-microtubule attachments.
  • Key proteins like Mad and Bub are involved, but their precise roles at kinetochores are unclear.
  • Mad1p and Mad2p form a complex, and Bub3p binds Bub1p, suggesting roles in kinetochore targeting.

Purpose of the Study:

  • To investigate the molecular interactions of spindle checkpoint proteins at kinetochores.
  • To elucidate the role of Mad1p, Bub1p, and Bub3p complex formation in checkpoint function.
  • To identify the regulatory mechanisms governing this complex during the cell cycle.

Main Methods:

  • Utilized budding yeast genetics to study protein interactions and complex formation.

Related Experiment Videos

  • Investigated protein complex levels under normal and activated spindle checkpoint conditions.
  • Employed site-directed mutagenesis to identify essential motifs for complex formation and checkpoint function.

    • Main Results:

    • Demonstrated a regulated association between Mad1p, Bub1p, and Bub3p in budding yeast.
    • Showed that this complex is upregulated upon spindle checkpoint activation.
    • Identified Mad2p and Mps1p as required for complex formation, while Mad3p and Bub2p are not.
    • Discovered a conserved motif in Mad1p critical for Mad1p-Bub1p-Bub3p complex assembly.

    Conclusions:

    • The Mad1p-Bub1p-Bub3p complex is a key component of the spindle checkpoint.
    • Formation of this complex, regulated by Mad2p and Mps1p, is essential for checkpoint activation.
    • A conserved motif in Mad1p is critical for the assembly of this complex and overall checkpoint efficacy.