Aurora B kinase cooperates with CENP-E to promote timely anaphase onset
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Error-free chromosome segregation requires that all chromosomes biorient on the mitotic spindle. The motor protein Centromere-associated protein E (CENP-E) facilitates chromosome congression by mediating the lateral sliding of sister chromatids along existing K-fibers, while the mitotic kinase Aurora B detaches kinetochore–microtubule interactions that are not bioriented. Whether these activities cooperate to promote efficient chromosome biorientation and timely anaphase onset is not known. We here show that the chromosomes that fail to congress after CENP-E depletion displayed high centromeric Aurora B kinase activity. This activity destabilized spindle pole proximal kinetochore–microtubule interactions resulting in a checkpoint-dependent mitotic delay that allowed CENP-E-independent chromosome congression, thus reducing chromosome segregation errors. This shows that Aurora B keeps the mitotic checkpoint active by destabilizing kinetochore fibers of polar chromosomes to permit chromosome congression in CENP-E-compromised cells and implies that this kinase normally prevents pole proximal syntelic attachments to allow CENP-E-mediated congression of mono-oriented chromosomes.
KeywordsElectronic Supplementary Material Nocodazole Spindle Pole Mitotic Checkpoint Supplementary Movie
We thank A. Musacchio, N. Galjart, and D. Cleveland for sharing the reagents and H. Maiato, M. Tanenbaum, G. Kops, and R. Medema for comments on the manuscript. A.F.M. is supported by a Ph.D. fellowship from Fundação para a Ciência e a Tecnologia (FCT) of Portugal and by a short-term fellowship from EMBO. This work was supported by grants from FCT (PTDC/BIA-BCM/64224/2006, C.E.S.) and the Netherlands Organization for Scientific Research (Vidi 917.66.332, S.M.A.L).
Animated time-lapse images of a control (siLuciferase) HeLa cell stably expressing YFP-H2B. Images were taken every 3 min. Overlay of DIC image (gray) and YFP-H2B (green), movie 1. YFP-H2B (gray), movie 2 (MPG 2112 kb)
Animated time-lapse images of a CENP-E-depleted HeLa cell stably expressing YFP-H2B. Phenotype a. Note that the cell only goes into anaphase after the last unaligned chromosome reaches the metaphase plate. Images were taken every 3 min. Overlay of DIC image (gray) and YFP-H2B (green), movie 3. YFP-H2B (gray), movie 4 (MPG 1940 kb)
Animated time-lapse images of a CENP-E-depleted HeLa cell stably expressing YFP-H2B. Phenotype b. Note that the cell goes into anaphase with an unaligned chromosome. Images were taken every 3 min. Overlay of DIC image (gray) and YFP-H2B (green), movie 5. YFP-H2B (gray), movie 6 (MPG 5566 kb)
Animated time-lapse images of a CENP-E-depleted HeLa cell stably expressing YFP-H2B. A black frame indicates that ZM447439 was added to the medium. Note that anaphase starts with several unaligned chromosomes. Images were taken every 3 min. Overlay of DIC image (gray) and YFP-H2B (green), movie 7. YFP-H2B (gray), movie 8 (MPG 3034 kb)
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