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PMID: 12221113 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Distinct chromosome segregation roles for spindle checkpoint proteins.

Molecular biology of the cell ·Vol. 13 ·No. 9 ·2002-09-00 ·Pages 3029-41

Warren CD, Brady DM, Johnston RC, Hanna JS, Hardwick KG, Spencer FA

Abstract

The spindle checkpoint plays a central role in the fidelity of chromosome transmission by ensuring that anaphase is initiated only after kinetochore-microtubule associations of all sister chromatid pairs are complete. In this study, we find that known spindle checkpoint proteins do not contribute equally to chromosome segregation fidelity in Saccharomyces cerevisiae. Loss of Bub1 or Bub3 protein elicits the largest effect. Analysis of Bub1p reveals the presence of two molecular functions. An N-terminal 608-amino acid (nonkinase) portion of the protein supports robust checkpoint activity, and, as expected, contributes to chromosome segregation. A C-terminal kinase-encoding segment independently contributes to chromosome segregation through an unknown mechanism. Both molecular functions depend on association with Bub3p. A 156-amino acid fragment of Bub1p functions in Bub3p binding and in kinetochore localization by one-hybrid assay. An adjacent segment is required for Mad1p binding, detected by deletion analysis and coimmunoprecipitation. Finally, overexpression of wild-type BUB1 or MAD3 genes leads to chromosome instability. Analysis of this activity indicates that the Bub3p-binding domain of Bub1p contributes to this phenotype through disruption of checkpoint activity as well as through introduction of kinetochore or spindle damage.

MeSH Terms
Alleles Cell Cycle Proteins/metabolism,physiology Chromosome Segregation Chromosomes/physiology,ultrastructure Fungal Proteins Immunoblotting Mutation Nuclear Proteins Phenotype Plasmids/metabolism Precipitin Tests Protein Binding Protein Kinases/physiology Protein Serine-Threonine Kinases Protein Structure, Tertiary Saccharomyces cerevisiae/genetics,physiology Saccharomyces cerevisiae Proteins Spindle Apparatus/physiology,ultrastructure Two-Hybrid System Techniques
Chemicals
BUB3 protein, S cerevisiae Cell Cycle Proteins Fungal Proteins MAD3 protein, S cerevisiae Nuclear Proteins Saccharomyces cerevisiae Proteins Protein Kinases Bub1 spindle checkpoint protein Protein Serine-Threonine Kinases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Warren Cheryl D
McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Brady D Michelle
Johnston Raymond C
Hanna Joseph S
Hardwick Kevin G
Spencer Forrest A
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e02-04-0203
Published
2002-09-00
Pages
3029-41
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC124140
Subset
IM
Grants
NIGMS NIH HHS · GM50842 · United States
External Links
PubMed source DOI: 10.1091/mbc.e02-04-0203 PMC Full Text View journal details
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