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PMID:14581449

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Citation

Westermann, S, Cheeseman, IM, Anderson, S, Yates, JR 3rd, Drubin, DG and Barnes, G (2003) Architecture of the budding yeast kinetochore reveals a conserved molecular core. J. Cell Biol. 163:215-22

Abstract

How kinetochore proteins are organized to connect chromosomes to spindle microtubules, and whether any structural and organizational themes are common to kinetochores from distantly related organisms, are key unanswered questions. Here, we used affinity chromatography and mass spectrometry to generate a map of kinetochore protein interactions. The budding yeast CENP-C homologue Mif2p specifically copurified with histones H2A, H2B, and H4, and with the histone H3-like CENP-A homologue Cse4p, strongly suggesting that Cse4p replaces histone H3 in a specialized centromeric nucleosome. A novel four-protein Mtw1 complex, the Nnf1p subunit of which has homology to the vertebrate kinetochore protein CENP-H, also copurified with Mif2p and a variety of central kinetochore proteins. We show that Mif2 is a critical in vivo target of the Aurora kinase Ipl1p. Chromatin immunoprecipitation studies demonstrated the biological relevance of these associations. We propose that a molecular core consisting of CENP-A, -C, -H, and Ndc80/HEC has been conserved from yeast to humans to link centromeres to spindle microtubules.

Links

PubMed PMC2173538 Online version:10.1083/jcb.200305100

Keywords

Amino Acid Sequence; Cell Cycle Proteins/chemistry; Cell Cycle Proteins/isolation & purification; Cell Cycle Proteins/metabolism; Centromere/metabolism; Chromatin/isolation & purification; Chromatin/metabolism; Chromosomal Proteins, Non-Histone/chemistry; Conserved Sequence; DNA-Binding Proteins/genetics; DNA-Binding Proteins/isolation & purification; DNA-Binding Proteins/metabolism; Fungal Proteins/chemistry; Histones/isolation & purification; Kinetochores/chemistry; Mitotic Spindle Apparatus/metabolism; Models, Biological; Molecular Sequence Data; Mutagenesis, Site-Directed; Nucleosomes/metabolism; Phosphorylation; Protein Subunits/chemistry; Protein Subunits/isolation & purification; Protein-Serine-Threonine Kinases/metabolism; Saccharomyces cerevisiae Proteins/genetics; Saccharomyces cerevisiae Proteins/isolation & purification; Saccharomyces cerevisiae Proteins/metabolism; Saccharomycetales/chemistry; Sequence Homology, Amino Acid

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

YEAST:CENPC

enables

GO:0044877: protein-containing complex binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

YEAST:IPL1

involved_in

GO:0006468: protein phosphorylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

  • has_input:(UniProtKB:P35201)|has_input:(UniProtKB:P40568)

Seeded From UniProt

complete

YEAST:IPL1

involved_in

GO:0006468: protein phosphorylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

has_input:(UniProtKB:P35201)|has_input:(UniProtKB:P40568)

Seeded From UniProt

complete


See also

References

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