The influence of catalysis on Mad2 activation dynamics

Marco Simonetta, Romilde Manzoni, Roberto Mosca, Marina Mapelli, Lucia Massimiliano, Martin Vink, Bela Novak, Andrea Musacchio, Andrea Ciliberto

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

Mad2 is a key component of the spindle assembly checkpoint, a safety device ensuring faithful sister chromatid separation in mitosis. The target of Mad2 is Cdc20, an activator of the anaphase-promoting complex/cyclosome (APC/ C). Mad2 binding to Cdc20 is a complex reaction that entails the conformational conversion of Mad2 from an open (O-Mad2) to a closed (C-Mad2) conformer. Previously, it has been hypothesized that the conversion of O-Mad2 is accelerated by its conformational dimerization with C-Mad2. This hypothesis, known as the Mad2-template hypothesis, is based on the unproven assumption that the natural conversion of O-Mad2 required to bind Cdc20 is slow. Here, we provide evidence for this fundamental assumption and demonstrate that conformational dimerization of Mad2 accelerates the rate of Mad2 binding to Cdc20. On the basis of our measurements, we developed a set of rate equations that deliver excellent predictions of experimental binding curves under a variety of different conditions. Our results strongly suggest that the interaction of Mad2 with Cdc20 is rate limiting for activation of the spindle checkpoint. Conformational dimerization of Mad2 is essential to accelerate Cdc20 binding, but it does not modify the equilibrium of the Mad2:Cdc20 interaction, i.e., it is purely catalytic. These results surpass previously formulated objections to the Mad2-template model and predict that the release of Mad2 from Cdc20 is an energy-driven process.

Original languageEnglish
Article numbere1000010
JournalPLoS Biology
Volume7
Issue number1
DOIs
Publication statusPublished - Jan 2009

Fingerprint

dimerization
Dimerization
Catalysis
catalytic activity
Chemical activation
safety equipment
Anaphase-Promoting Complex-Cyclosome
Protective Devices
M Phase Cell Cycle Checkpoints
Safety devices
process energy
Chromatids
chromatids
Mitosis
mitosis
prediction

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

Simonetta, M., Manzoni, R., Mosca, R., Mapelli, M., Massimiliano, L., Vink, M., ... Ciliberto, A. (2009). The influence of catalysis on Mad2 activation dynamics. PLoS Biology, 7(1), [e1000010]. https://doi.org/10.1371/journal.pbio.1000010

The influence of catalysis on Mad2 activation dynamics. / Simonetta, Marco; Manzoni, Romilde; Mosca, Roberto; Mapelli, Marina; Massimiliano, Lucia; Vink, Martin; Novak, Bela; Musacchio, Andrea; Ciliberto, Andrea.

In: PLoS Biology, Vol. 7, No. 1, e1000010, 01.2009.

Research output: Contribution to journalArticle

Simonetta, M, Manzoni, R, Mosca, R, Mapelli, M, Massimiliano, L, Vink, M, Novak, B, Musacchio, A & Ciliberto, A 2009, 'The influence of catalysis on Mad2 activation dynamics', PLoS Biology, vol. 7, no. 1, e1000010. https://doi.org/10.1371/journal.pbio.1000010
Simonetta, Marco ; Manzoni, Romilde ; Mosca, Roberto ; Mapelli, Marina ; Massimiliano, Lucia ; Vink, Martin ; Novak, Bela ; Musacchio, Andrea ; Ciliberto, Andrea. / The influence of catalysis on Mad2 activation dynamics. In: PLoS Biology. 2009 ; Vol. 7, No. 1.
@article{3538b4ef1d5340c7a411b7343d22984d,
title = "The influence of catalysis on Mad2 activation dynamics",
abstract = "Mad2 is a key component of the spindle assembly checkpoint, a safety device ensuring faithful sister chromatid separation in mitosis. The target of Mad2 is Cdc20, an activator of the anaphase-promoting complex/cyclosome (APC/ C). Mad2 binding to Cdc20 is a complex reaction that entails the conformational conversion of Mad2 from an open (O-Mad2) to a closed (C-Mad2) conformer. Previously, it has been hypothesized that the conversion of O-Mad2 is accelerated by its conformational dimerization with C-Mad2. This hypothesis, known as the Mad2-template hypothesis, is based on the unproven assumption that the natural conversion of O-Mad2 required to bind Cdc20 is slow. Here, we provide evidence for this fundamental assumption and demonstrate that conformational dimerization of Mad2 accelerates the rate of Mad2 binding to Cdc20. On the basis of our measurements, we developed a set of rate equations that deliver excellent predictions of experimental binding curves under a variety of different conditions. Our results strongly suggest that the interaction of Mad2 with Cdc20 is rate limiting for activation of the spindle checkpoint. Conformational dimerization of Mad2 is essential to accelerate Cdc20 binding, but it does not modify the equilibrium of the Mad2:Cdc20 interaction, i.e., it is purely catalytic. These results surpass previously formulated objections to the Mad2-template model and predict that the release of Mad2 from Cdc20 is an energy-driven process.",
author = "Marco Simonetta and Romilde Manzoni and Roberto Mosca and Marina Mapelli and Lucia Massimiliano and Martin Vink and Bela Novak and Andrea Musacchio and Andrea Ciliberto",
year = "2009",
month = "1",
doi = "10.1371/journal.pbio.1000010",
language = "English",
volume = "7",
journal = "PLoS Biology",
issn = "1544-9173",
publisher = "Public Library of Science",
number = "1",

}

TY - JOUR

T1 - The influence of catalysis on Mad2 activation dynamics

AU - Simonetta, Marco

AU - Manzoni, Romilde

AU - Mosca, Roberto

AU - Mapelli, Marina

AU - Massimiliano, Lucia

AU - Vink, Martin

AU - Novak, Bela

AU - Musacchio, Andrea

AU - Ciliberto, Andrea

PY - 2009/1

Y1 - 2009/1

N2 - Mad2 is a key component of the spindle assembly checkpoint, a safety device ensuring faithful sister chromatid separation in mitosis. The target of Mad2 is Cdc20, an activator of the anaphase-promoting complex/cyclosome (APC/ C). Mad2 binding to Cdc20 is a complex reaction that entails the conformational conversion of Mad2 from an open (O-Mad2) to a closed (C-Mad2) conformer. Previously, it has been hypothesized that the conversion of O-Mad2 is accelerated by its conformational dimerization with C-Mad2. This hypothesis, known as the Mad2-template hypothesis, is based on the unproven assumption that the natural conversion of O-Mad2 required to bind Cdc20 is slow. Here, we provide evidence for this fundamental assumption and demonstrate that conformational dimerization of Mad2 accelerates the rate of Mad2 binding to Cdc20. On the basis of our measurements, we developed a set of rate equations that deliver excellent predictions of experimental binding curves under a variety of different conditions. Our results strongly suggest that the interaction of Mad2 with Cdc20 is rate limiting for activation of the spindle checkpoint. Conformational dimerization of Mad2 is essential to accelerate Cdc20 binding, but it does not modify the equilibrium of the Mad2:Cdc20 interaction, i.e., it is purely catalytic. These results surpass previously formulated objections to the Mad2-template model and predict that the release of Mad2 from Cdc20 is an energy-driven process.

AB - Mad2 is a key component of the spindle assembly checkpoint, a safety device ensuring faithful sister chromatid separation in mitosis. The target of Mad2 is Cdc20, an activator of the anaphase-promoting complex/cyclosome (APC/ C). Mad2 binding to Cdc20 is a complex reaction that entails the conformational conversion of Mad2 from an open (O-Mad2) to a closed (C-Mad2) conformer. Previously, it has been hypothesized that the conversion of O-Mad2 is accelerated by its conformational dimerization with C-Mad2. This hypothesis, known as the Mad2-template hypothesis, is based on the unproven assumption that the natural conversion of O-Mad2 required to bind Cdc20 is slow. Here, we provide evidence for this fundamental assumption and demonstrate that conformational dimerization of Mad2 accelerates the rate of Mad2 binding to Cdc20. On the basis of our measurements, we developed a set of rate equations that deliver excellent predictions of experimental binding curves under a variety of different conditions. Our results strongly suggest that the interaction of Mad2 with Cdc20 is rate limiting for activation of the spindle checkpoint. Conformational dimerization of Mad2 is essential to accelerate Cdc20 binding, but it does not modify the equilibrium of the Mad2:Cdc20 interaction, i.e., it is purely catalytic. These results surpass previously formulated objections to the Mad2-template model and predict that the release of Mad2 from Cdc20 is an energy-driven process.

UR - http://www.scopus.com/inward/record.url?scp=58849147890&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=58849147890&partnerID=8YFLogxK

U2 - 10.1371/journal.pbio.1000010

DO - 10.1371/journal.pbio.1000010

M3 - Article

VL - 7

JO - PLoS Biology

JF - PLoS Biology

SN - 1544-9173

IS - 1

M1 - e1000010

ER -