Retention of the Bub3 checkpoint protein on lagging chromosomes

Maria J. Martinez-Exposito, Kenneth B. Kaplan, Jay Copeland, Peter K. Sorger

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Accurate chromosome segregation at mitosis is ensured both by the intrinsic fidelity of the mitotic machinery and by the operation of checkpoints that monitor chromosome-microtubule attachment. When unattached kinetochores are present, anaphase is delayed and the time available for chromosome-microtubule capture increases. Genes required for this delay first were identified in budding yeast (the MAD and BUB genes), but it is not yet known how the checkpoint senses unattached chromosomes or how it signals cell-cycle arrest. We report the isolation and analysis of a murine homologue of BUB3, a gene whose deletion abolishes mitotic checkpoint function in Saccharomyces cerevisiae, mBub3 belongs to a small gene family that has been highly conserved through evolution. By expressing recombinant proteins in insect cells, we show that mBub3, like yeast Bub3p, binds to Bub1 to form a complex with protein kinase activity. During prophase and prometaphase, preceding kinetochore-microtubule attachment, Bub3 localizes to kinetochores. High levels of mBub3 remain associated with lagging chromosomes but not with correctly aligned chromosomes during metaphase, consistent with a role for Bub3 in sensing microtubule attachment. Intriguingly, the number of lagging chromosomes with high Bub3 staining increases dramatically in cells treated with low (and pharmacologically relevant) concentrations of the chemotherapeutic taxol and the microtubule poison nocodazole.

Original languageEnglish (US)
Pages (from-to)8493-8498
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number15
DOIs
StatePublished - Jul 20 1999
Externally publishedYes

Fingerprint

Microtubules
Chromosomes
Kinetochores
Proteins
Prometaphase
M Phase Cell Cycle Checkpoints
Genes
Nocodazole
Prophase
Chromosome Segregation
Anaphase
Saccharomycetales
Poisons
Gene Deletion
Metaphase
Paclitaxel
Cell Cycle Checkpoints
Recombinant Proteins
Mitosis
Protein Kinases

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Retention of the Bub3 checkpoint protein on lagging chromosomes. / Martinez-Exposito, Maria J.; Kaplan, Kenneth B.; Copeland, Jay; Sorger, Peter K.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, No. 15, 20.07.1999, p. 8493-8498.

Research output: Contribution to journalArticle

Martinez-Exposito, Maria J. ; Kaplan, Kenneth B. ; Copeland, Jay ; Sorger, Peter K. / Retention of the Bub3 checkpoint protein on lagging chromosomes. In: Proceedings of the National Academy of Sciences of the United States of America. 1999 ; Vol. 96, No. 15. pp. 8493-8498.
@article{75ac8070b72649f3b915137af9b564b3,
title = "Retention of the Bub3 checkpoint protein on lagging chromosomes",
abstract = "Accurate chromosome segregation at mitosis is ensured both by the intrinsic fidelity of the mitotic machinery and by the operation of checkpoints that monitor chromosome-microtubule attachment. When unattached kinetochores are present, anaphase is delayed and the time available for chromosome-microtubule capture increases. Genes required for this delay first were identified in budding yeast (the MAD and BUB genes), but it is not yet known how the checkpoint senses unattached chromosomes or how it signals cell-cycle arrest. We report the isolation and analysis of a murine homologue of BUB3, a gene whose deletion abolishes mitotic checkpoint function in Saccharomyces cerevisiae, mBub3 belongs to a small gene family that has been highly conserved through evolution. By expressing recombinant proteins in insect cells, we show that mBub3, like yeast Bub3p, binds to Bub1 to form a complex with protein kinase activity. During prophase and prometaphase, preceding kinetochore-microtubule attachment, Bub3 localizes to kinetochores. High levels of mBub3 remain associated with lagging chromosomes but not with correctly aligned chromosomes during metaphase, consistent with a role for Bub3 in sensing microtubule attachment. Intriguingly, the number of lagging chromosomes with high Bub3 staining increases dramatically in cells treated with low (and pharmacologically relevant) concentrations of the chemotherapeutic taxol and the microtubule poison nocodazole.",
author = "Martinez-Exposito, {Maria J.} and Kaplan, {Kenneth B.} and Jay Copeland and Sorger, {Peter K.}",
year = "1999",
month = "7",
day = "20",
doi = "10.1073/pnas.96.15.8493",
language = "English (US)",
volume = "96",
pages = "8493--8498",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "15",

}

TY - JOUR

T1 - Retention of the Bub3 checkpoint protein on lagging chromosomes

AU - Martinez-Exposito, Maria J.

AU - Kaplan, Kenneth B.

AU - Copeland, Jay

AU - Sorger, Peter K.

PY - 1999/7/20

Y1 - 1999/7/20

N2 - Accurate chromosome segregation at mitosis is ensured both by the intrinsic fidelity of the mitotic machinery and by the operation of checkpoints that monitor chromosome-microtubule attachment. When unattached kinetochores are present, anaphase is delayed and the time available for chromosome-microtubule capture increases. Genes required for this delay first were identified in budding yeast (the MAD and BUB genes), but it is not yet known how the checkpoint senses unattached chromosomes or how it signals cell-cycle arrest. We report the isolation and analysis of a murine homologue of BUB3, a gene whose deletion abolishes mitotic checkpoint function in Saccharomyces cerevisiae, mBub3 belongs to a small gene family that has been highly conserved through evolution. By expressing recombinant proteins in insect cells, we show that mBub3, like yeast Bub3p, binds to Bub1 to form a complex with protein kinase activity. During prophase and prometaphase, preceding kinetochore-microtubule attachment, Bub3 localizes to kinetochores. High levels of mBub3 remain associated with lagging chromosomes but not with correctly aligned chromosomes during metaphase, consistent with a role for Bub3 in sensing microtubule attachment. Intriguingly, the number of lagging chromosomes with high Bub3 staining increases dramatically in cells treated with low (and pharmacologically relevant) concentrations of the chemotherapeutic taxol and the microtubule poison nocodazole.

AB - Accurate chromosome segregation at mitosis is ensured both by the intrinsic fidelity of the mitotic machinery and by the operation of checkpoints that monitor chromosome-microtubule attachment. When unattached kinetochores are present, anaphase is delayed and the time available for chromosome-microtubule capture increases. Genes required for this delay first were identified in budding yeast (the MAD and BUB genes), but it is not yet known how the checkpoint senses unattached chromosomes or how it signals cell-cycle arrest. We report the isolation and analysis of a murine homologue of BUB3, a gene whose deletion abolishes mitotic checkpoint function in Saccharomyces cerevisiae, mBub3 belongs to a small gene family that has been highly conserved through evolution. By expressing recombinant proteins in insect cells, we show that mBub3, like yeast Bub3p, binds to Bub1 to form a complex with protein kinase activity. During prophase and prometaphase, preceding kinetochore-microtubule attachment, Bub3 localizes to kinetochores. High levels of mBub3 remain associated with lagging chromosomes but not with correctly aligned chromosomes during metaphase, consistent with a role for Bub3 in sensing microtubule attachment. Intriguingly, the number of lagging chromosomes with high Bub3 staining increases dramatically in cells treated with low (and pharmacologically relevant) concentrations of the chemotherapeutic taxol and the microtubule poison nocodazole.

UR - http://www.scopus.com/inward/record.url?scp=0033587725&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033587725&partnerID=8YFLogxK

U2 - 10.1073/pnas.96.15.8493

DO - 10.1073/pnas.96.15.8493

M3 - Article

C2 - 10411903

AN - SCOPUS:0033587725

VL - 96

SP - 8493

EP - 8498

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 15

ER -