Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
???displayArticle.abstract???
We show that the Xenopus homologs of Ndc80/Tid3/HEC1 (xNdc80) and Nuf2/MPP1/Him-10 (xNuf2) proteins physically interact in a 190-kD complex that associates with the outer kinetochore from prometaphase through anaphase. Injecting function-blocking antibodies to either xNdc80 or xNuf2 into XTC cells caused premature exit from mitosis without detectable chromosome congression or anaphase movements. Injected cells did not arrest in response to microtubule drugs, showing that the complex is required for the spindle checkpoint. Kinetochores assembled in Xenopus extracts after immunodepletion of the complex did not contain xRod, xZw10, xP150 glued (Dynactin), xMad1, xMad2, xBub1, and xBub3, demonstrating that the xNdc80 complex is required for functional kinetochore assembly. In contrast, function-blocking antibodies did not affect the localization of other kinetochore proteins when added to extracts containing previously assembled kinetochores. These extracts with intact kinetochores were deficient in checkpoint signaling, suggesting that the Ndc80 complex participates in the spindle checkpoint. We also demonstrate that the spindle checkpoint can arrest budding yeast cells lacking Ndc80 or Nuf2, whereas yeast lacking both proteins fail to arrest in mitosis. Systematic deletion of yeast kinetochore genes suggests that the Ndc80 complex has a unique role in spindle checkpoint signaling. We propose that the Ndc80 complex has conserved roles in kinetochore assembly, chromosome congression, and spindle checkpoint signaling.
???displayArticle.pubmedLink???
12514103 ???displayArticle.pmcLink???PMC195965 ???displayArticle.link???Genes Dev ???displayArticle.grants???[+]
Adams,
Relationship of actin and tubulin distribution to bud growth in wild-type and morphogenetic-mutant Saccharomyces cerevisiae.
1984, Pubmed
Adams,
Relationship of actin and tubulin distribution to bud growth in wild-type and morphogenetic-mutant Saccharomyces cerevisiae.
1984,
Pubmed Basto,
Rough deal and Zw10 are required for the metaphase checkpoint in Drosophila.
2000,
Pubmed Bolton,
Aurora B kinase exists in a complex with survivin and INCENP and its kinase activity is stimulated by survivin binding and phosphorylation.
2002,
Pubmed
,
Xenbase Brown,
MIF2 is required for mitotic spindle integrity during anaphase spindle elongation in Saccharomyces cerevisiae.
1993,
Pubmed Chan,
Human Zw10 and ROD are mitotic checkpoint proteins that bind to kinetochores.
2000,
Pubmed Cheeseman,
Mitotic spindle integrity and kinetochore function linked by the Duo1p/Dam1p complex.
2001,
Pubmed Cheeseman,
Simple centromere, complex kinetochore: linking spindle microtubules and centromeric DNA in budding yeast.
2002,
Pubmed Chen,
HEC, a novel nuclear protein rich in leucine heptad repeats specifically involved in mitosis.
1997,
Pubmed Chen,
Spindle checkpoint protein Xmad1 recruits Xmad2 to unattached kinetochores.
1998,
Pubmed
,
Xenbase Cooke,
The inner centromere protein (INCENP) antigens: movement from inner centromere to midbody during mitosis.
1987,
Pubmed Dohmen,
Heat-inducible degron: a method for constructing temperature-sensitive mutants.
1994,
Pubmed Durfee,
The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit.
1993,
Pubmed Earnshaw,
Identification of a family of human centromere proteins using autoimmune sera from patients with scleroderma.
1985,
Pubmed Gardner,
The spindle checkpoint of the yeast Saccharomyces cerevisiae requires kinetochore function and maps to the CBF3 domain.
2001,
Pubmed Gardner,
The spindle checkpoint: two transitions, two pathways.
2000,
Pubmed Hartwell,
Macromolecule synthesis in temperature-sensitive mutants of yeast.
1967,
Pubmed He,
Molecular analysis of kinetochore-microtubule attachment in budding yeast.
2001,
Pubmed Howe,
HIM-10 is required for kinetochore structure and function on Caenorhabditis elegans holocentric chromosomes.
2001,
Pubmed Howell,
Visualization of Mad2 dynamics at kinetochores, along spindle fibers, and at spindle poles in living cells.
2000,
Pubmed Hoyt,
S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function.
1991,
Pubmed Janke,
Four new subunits of the Dam1-Duo1 complex reveal novel functions in sister kinetochore biorientation.
2002,
Pubmed Janke,
The budding yeast proteins Spc24p and Spc25p interact with Ndc80p and Nuf2p at the kinetochore and are important for kinetochore clustering and checkpoint control.
2001,
Pubmed Jones,
Yeast Dam1p is required to maintain spindle integrity during mitosis and interacts with the Mps1p kinase.
1999,
Pubmed Kallio,
Rapid microtubule-independent dynamics of Cdc20 at kinetochores and centrosomes in mammalian cells.
2002,
Pubmed King,
How proteolysis drives the cell cycle.
1996,
Pubmed Li,
The mitotic spindle is required for loading of the DASH complex onto the kinetochore.
2002,
Pubmed Li,
Feedback control of mitosis in budding yeast.
1991,
Pubmed Lupas,
Predicting coiled coils from protein sequences.
1991,
Pubmed Martin-Lluesma,
Role of Hec1 in spindle checkpoint signaling and kinetochore recruitment of Mad1/Mad2.
2002,
Pubmed Minshull,
A MAP kinase-dependent spindle assembly checkpoint in Xenopus egg extracts.
1994,
Pubmed
,
Xenbase Murray,
Cell cycle extracts.
1991,
Pubmed Nabetani,
A conserved protein, Nuf2, is implicated in connecting the centromere to the spindle during chromosome segregation: a link between the kinetochore function and the spindle checkpoint.
2001,
Pubmed Newman,
A computationally directed screen identifying interacting coiled coils from Saccharomyces cerevisiae.
2000,
Pubmed Osborne,
Nuf2, a spindle pole body-associated protein required for nuclear division in yeast.
1994,
Pubmed Pereira,
The Bub2p spindle checkpoint links nuclear migration with mitotic exit.
2000,
Pubmed Saitoh,
CENP-C, an autoantigen in scleroderma, is a component of the human inner kinetochore plate.
1992,
Pubmed Severin,
Stu2 promotes mitotic spindle elongation in anaphase.
2001,
Pubmed Shah,
Waiting for anaphase: Mad2 and the spindle assembly checkpoint.
2000,
Pubmed Siegel,
Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases.
1966,
Pubmed Stukenberg,
Systematic identification of mitotic phosphoproteins.
1997,
Pubmed
,
Xenbase Wang,
The Bfa1/Bub2 GAP complex comprises a universal checkpoint required to prevent mitotic exit.
2000,
Pubmed Wigge,
Analysis of the Saccharomyces spindle pole by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry.
1998,
Pubmed Wigge,
The Ndc80p complex from Saccharomyces cerevisiae contains conserved centromere components and has a function in chromosome segregation.
2001,
Pubmed Winey,
MPS1 and MPS2: novel yeast genes defining distinct steps of spindle pole body duplication.
1991,
Pubmed Wordeman,
Chemical subdomains within the kinetochore domain of isolated CHO mitotic chromosomes.
1991,
Pubmed
