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Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast
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BuGZ is required for Bub3 stability, Bub1 kinetochore function, and chromosome alignment.

Chad M Toledo1, Jacob A Herman2, Jonathan B Olsen3

  • 1Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA; Molecular and Cellular Biology Program, University of Washington, Seattle, WA 98195, USA.

Developmental Cell
|January 28, 2014
PubMed
Summary
This summary is machine-generated.

A novel spindle assembly checkpoint (SAC) protein, BuGZ, stabilizes Bub3 and is crucial for chromosome alignment in cancer cells. Its absence causes lethal defects, highlighting its role in higher eukaryotes.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Cancer Research

Background:

  • The spindle assembly checkpoint (SAC) ensures accurate chromosome segregation during mitosis.
  • Proper kinetochore-microtubule attachment and tension are monitored by the SAC to prevent premature anaphase onset.
  • Defects in SAC function are implicated in aneuploidy and cancer development.

Purpose of the Study:

  • To identify novel components of the spindle assembly checkpoint (SAC).
  • To investigate the function of BuGZ/ZNF207 in mitosis and chromosome congression.
  • To understand the role of BuGZ in cancer cell viability.

Main Methods:

  • RNA interference (RNAi) viability screen in human glioblastoma multiforme (GBM) cells.
  • Co-immunoprecipitation to assess protein binding and stabilization.
  • Immunofluorescence microscopy to analyze protein localization at kinetochores (KTs).
  • Biochemical assays to measure kinase activity and protein phosphorylation.
  • Phylogenetic analysis of BuGZ orthologs.

Main Results:

  • BuGZ/ZNF207 was identified as a novel SAC component from a screen in GBM stem cells.
  • BuGZ binds to and stabilizes Bub3 via its GLE2p-binding sequence (GLEBS) domain.
  • BuGZ inhibition leads to loss of Bub3 and Bub1 from KTs, reduced H2A phosphorylation, and decreased Aurora B kinase activity.
  • BuGZ depletion causes lethal chromosome congression defects in cancer cells.
  • BuGZ orthologs are conserved in higher eukaryotes but absent in yeasts.

Conclusions:

  • BuGZ is essential for maintaining Bub3 stability and proper kinetochore function during mitosis.
  • BuGZ plays a critical role in chromosome alignment and is vital for cancer cell survival.
  • The evolution of BuGZ likely contributed to robust chromosome segregation mechanisms in higher eukaryotes.