Structural analysis of Bub3 interactions in the mitotic spindle checkpoint

Nicholas A. Larsen, Jawdat Al-Bassam, Ronnie R. Wei, Stephen C. Harrison

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

The Mad3/BubR1, Mad2, Bub1, and Bub3 proteins are gatekeepers for the transition from metaphase to anaphase. Mad3 from Saccharomyces cerevisiae has homology to Bub1 but lacks a corresponding C-terminal kinase domain. Mad3 forms a stable heterodimer with Bub3. Negative-stain electron microscopy shows that Mad3 is an extended molecule (≈200 Å long), whereas Bub3 is globular. The Gle2-binding-sequence (GLEBS) motifs found in Mad3 and Bub1 are necessary and sufficient for interaction with Bub3. The calorimetrically determined dissociation constants for GLEBS-motif peptides and BubS are ≈5 μM. Crystal structures of these peptides with Bub3 show that the interactions for Mad3 and Bub1 are similar and mutually exclusive. In both structures, the GLEBS peptide snakes along the top surface of the β-propeller, forming an extensive interface. Mutations in either protein that disrupt the interface cause checkpoint deficiency and chromosome instability. We propose that the structure imposed on the GLEBS segment by its association with Bub3 enables recruitment to unattached kinetochores.

Original languageEnglish (US)
Pages (from-to)1201-1206
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume104
Issue number4
DOIs
StatePublished - Jan 23 2007
Externally publishedYes

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M Phase Cell Cycle Checkpoints
Peptides
Kinetochores
Chromosomal Instability
Anaphase
Snakes
Metaphase
Saccharomyces cerevisiae
Electron Microscopy
Proteins
Coloring Agents
Phosphotransferases
Mutation

Keywords

  • β-propeller
  • Crystal structures
  • GLEBS motif
  • Rae1/Nup98

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Structural analysis of Bub3 interactions in the mitotic spindle checkpoint. / Larsen, Nicholas A.; Al-Bassam, Jawdat; Wei, Ronnie R.; Harrison, Stephen C.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, No. 4, 23.01.2007, p. 1201-1206.

Research output: Contribution to journalArticle

Larsen, Nicholas A. ; Al-Bassam, Jawdat ; Wei, Ronnie R. ; Harrison, Stephen C. / Structural analysis of Bub3 interactions in the mitotic spindle checkpoint. In: Proceedings of the National Academy of Sciences of the United States of America. 2007 ; Vol. 104, No. 4. pp. 1201-1206.
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