Accumulation of Mad2-Cdc20 complex during spindle checkpoint activation requires binding of open and closed conformers of Mad2 in Saccharomyces cerevisiae

Luigi Nezi, Giulia Rancati, Anna De Antoni, Sebastiano Pasqualato, Simonetta Piatti, Andrea Musacchio

Research output: Contribution to journalArticle

Abstract

The spindle assembly checkpoint (SAC) coordinates mitotic progression with sister chromatid alignment. In mitosis, the checkpoint machinery accumulates at kinetochores, which are scaffolds devoted to microtubule capture. The checkpoint protein Mad2 (mitotic arrest de-. cient 2) adopts two conformations: open (O-Mad2) and closed (C-Mad2). C-Mad2 forms when Mad2 binds its checkpoint target Cdc20 or its kinetochore receptor Mad1. When unbound to these ligands, Mad2 folds as O-Mad2. In HeLa cells, an essential interaction between C- and O-Mad2 conformers allows Mad1-bound C-Mad2 to recruit cytosolic O-Mad2 to kinetochores. In this study, we show that the interaction of the O and C conformers of Mad2 is conserved in Saccharomyces cerevisiae. MAD2 mutant alleles impaired in this interaction fail to restore the SAC in a mad2 deletion strain. The corresponding mutant proteins bind Mad1 normally, but their ability to bind Cdc20 is dramatically impaired in vivo. Our biochemical and genetic evidence shows that the interaction of O- and C-Mad2 is essential for the SAC and is conserved in evolution.

Original languageEnglish
Pages (from-to)39-51
Number of pages13
JournalJournal of Cell Biology
Volume174
Issue number1
DOIs
Publication statusPublished - Jul 3 2006

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M Phase Cell Cycle Checkpoints
Kinetochores
Saccharomyces cerevisiae
Chromatids
Mutant Proteins
HeLa Cells
Mitosis
Cell Communication
Microtubules
Molecular Biology
Alleles
Ligands
Proteins

ASJC Scopus subject areas

  • Cell Biology

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Accumulation of Mad2-Cdc20 complex during spindle checkpoint activation requires binding of open and closed conformers of Mad2 in Saccharomyces cerevisiae. / Nezi, Luigi; Rancati, Giulia; De Antoni, Anna; Pasqualato, Sebastiano; Piatti, Simonetta; Musacchio, Andrea.

In: Journal of Cell Biology, Vol. 174, No. 1, 03.07.2006, p. 39-51.

Research output: Contribution to journalArticle

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