title: Centromere-associated protein-E is essential for the mammalian mitotic checkpoint to prevent aneuploidy due to single chromosome loss.
abstract: Centromere-associated protein-E (CENP-E) is an essential mitotic kinesin that is required for efficient, stable microtubule capture at kinetochores. It also directly binds to BubR1, a kinetochore-associated kinase implicated in the mitotic checkpoint, the major cell cycle control pathway in which unattached kinetochores prevent anaphase onset. Here, we show that single unattached kinetochores depleted of CENP-E cannot block entry into anaphase, resulting in aneuploidy in 25% of divisions in primary mouse fibroblasts in vitro and in 95% of regenerating hepatocytes in vivo. Without CENP-E, diminished levels of BubR1 are recruited to kinetochores and BubR1 kinase activity remains at basal levels. CENP-E binds to and directly stimulates the kinase activity of purified BubR1 in vitro. Thus, CENP-E is required for enhancing recruitment of its binding partner BubR1 to each unattached kinetochore and for stimulating BubR1 kinase activity, implicating it as an essential amplifier of a basal mitotic checkpoint signal.
authors.affiliation: Ludwig Institute for Cancer Research, 3080 CMM-East, 9500 Gilman Drive, La Jolla, CA 92093-0670, USA.
authors.forename: Beth A A
authors.initials: BA
authors.lastname: Weaver
authors.forename: Zahid Q
authors.initials: ZQ
authors.lastname: Bonday
authors.forename: Frances R
authors.initials: FR
authors.lastname: Putkey
authors.forename: Geert J P L
authors.initials: GJ
authors.lastname: Kops
authors.forename: Alain D
authors.initials: AD
authors.lastname: Silk
authors.forename: Don W
authors.initials: DW
authors.lastname: Cleveland
mesh_terms: D000782:Aneuploidy; D000818:Animals; D018797:Cell Cycle Proteins; D002503:Centromere / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D005347:Fibroblasts; D006367:HeLa Cells; D006801:Humans; D019426:Integrases / Q000378:metabolism; D018386:Kinetochores / Q000378:metabolism; D051379:Mice; D008938:Mitosis / Q000502:physiology*; D011494:Protein Kinases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000378:metabolism
chemical_list: C584278:Bub1b protein, mouse; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D014764:Viral Proteins; C068688:centromere protein E; D011494:Protein Kinases; C584277:BUB1 protein, human; C090464:Bub1 spindle checkpoint protein; D017346:Protein Serine-Threonine Kinases; C045073:Cre recombinase; D019426:Integrases