title: Kinetochore stretching inactivates the spindle assembly checkpoint.
abstract: The spindle assembly checkpoint (SAC) monitors the attachment of microtubules to the kinetochore and inhibits anaphase when microtubule binding is incomplete. The SAC might also respond to tension; however, how cells can sense tension and whether its detection is important to satisfy the SAC remain controversial. We generated a HeLa cell line in which two components of the kinetochore, centromere protein A and Mis12, are labeled with green and red fluorophores, respectively. Live cell imaging of these cells reveals repetitive cycles of kinetochore extension and recoiling after biorientation. Under conditions in which kinetochore stretching is suppressed, cells fail to silence the SAC and enter anaphase after a delay, regardless of centromere stretching. Monitoring cyclin B levels as a readout for anaphase-promoting complex/cyclosome activity, we find that suppression of kinetochore stretching delays and decelerates cyclin B degradation. These observations suggest that the SAC monitors stretching of kinetochores rather than centromeres and that kinetochore stretching promotes silencing of the SAC signal.
authors.affiliation: Cancer Institute, Japanese Foundation for Cancer Research, Koto-ku, Tokyo 135-8550, Japan.
authors.forename: Kazuhiko S K
authors.initials: KS
authors.lastname: Uchida
authors.forename: Kentaro
authors.initials: K
authors.lastname: Takagaki
authors.forename: Kazuki
authors.initials: K
authors.lastname: Kumada
authors.forename: Youko
authors.initials: Y
authors.lastname: Hirayama
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Noda
authors.forename: Toru
authors.initials: T
authors.lastname: Hirota
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D001324:Autoantigens / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology*; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism; D015739:Nocodazole / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism*; D013314:Stress, Mechanical; D050257:Tubulin Modulators / Q000378:metabolism
chemical_list: D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; C492664:MIS12 protein, human; D008869:Microtubule-Associated Proteins; D046912:Multiprotein Complexes; D011993:Recombinant Fusion Proteins; D050257:Tubulin Modulators; C108312:condensin complexes; D000251:Adenosine Triphosphatases; D015739:Nocodazole