The inhibition of Aurora A abrogates the mitotic delay induced by microtubule perturbing agents.
The spindle assembly checkpoint functions during mitosis to ensure that chromosomes are properly aligned in mitotic cells prior to the onset of anaphase, thereby ensuring an equal segregation of genetic material to each daughter cell. Defects in the function of this checkpoint lead to aneuploidy, and eventually to cell death or senescence. The Aurora-related kinases, and in particular Aurora B, have been shown to play a role in regulating the spindle assembly checkpoint. In this study, we demonstrate that Aurora A activity is required for maintainance of the spindle assembly checkpoint mediated-mitotic delay induced by microtubule perturbing agents. Inhibition of Aurora A using MLN8054, a selective small-molecule inhibitor of Aurora A, in paclitaxel- or nocodazole-treated cells induces cells to become multinucleated. Using time-lapse microscopy, we demonstrate that the multinucleation phenotype arises via mitotic slippage, which is significantly accelerated upon Aurora A inhibition. Under these conditions, the spindle assembly checkpoint protein BubR1 remains localized to kinetochores prior to mitotic slippage. Moreover, we demonstrate that Aurora B remains active in these mitotic cells, indicating that the mitotic slippage induced by MLN8054 is most likely due to the inhibition of Aurora A. This finding was corroborated by demonstrating that Aurora A depletion using RNA interference in paclitaxel-treated cells also induces multinucleation. Taken together, these results suggest that Aurora A is necessary for the maintenance of the mitotic delay induced in response to microtubule-perturbing agents.
Wysong
Deborah R
DR
Department of Molecular and Cellular Oncology, Millennium Pharmaceuticals, Inc, Cambridge, Massachusetts, USA.
Chakravarty
Arijit
A
Hoar
Kara
K
Ecsedy
Jeffrey A
JA
D064107:Aurora Kinase B; D064127:Aurora Kinases; D001552:Benzazepines / Q000494:pharmacology; D045744:Cell Line, Tumor; D018816:Genes, cdc / Q000187:drug effects / Q000502:physiology; D006801:Humans; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D008938:Mitosis / Q000187:drug effects; D015739:Nocodazole / Q000494:pharmacology; D017239:Paclitaxel / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D034741:RNA, Small Interfering / Q000378:metabolism; D008941:Spindle Apparatus / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D050257:Tubulin Modulators / Q000494:pharmacology
D001552:Benzazepines; C518940:MLN8054; D034741:RNA, Small Interfering; D050257:Tubulin Modulators; C576921:AURKB protein, human; D064107:Aurora Kinase B; D064127:Aurora Kinases; C584277:BUB1 protein, human; C090464:Bub1 spindle checkpoint protein; D017346:Protein Serine-Threonine Kinases; D017239:Paclitaxel; D015739:Nocodazole