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Survivin is required for stable checkpoint activation in taxol-treated HeLa cells
journal contribution
posted on 2023-06-08, 09:37 authored by Ana Carvalho, Mar Carmena, Clara Sambade, William C Earnshaw, Sally P WheatleySurvivin is an essential chromosomal passenger protein whose function remains unclear. Here, we have used RNA interference to specifically repress Survivin in cultured HeLa cells. Immunoblot analysis showed that Survivin was no longer detectable in cultures 60 hours after transfection with Survivin-specific siRNA. Live cell analysis showed that many Survivin-depleted cells were delayed in mitosis, and immunofluorescence analysis of fixed specimens revealed that Survivin-depleted cells accumulated in prometaphase with misaligned chromosomes. The chromosomal passenger proteins, INCENP and Aurora-B, which can interact directly with Survivin, were absent from the centromeres of Survivin-depleted cells. These data contribute to the emerging picture that Survivin operates together with INCENP and Aurora-B to perform its mitotic duties. Some Survivin-depleted cells eventually exited mitosis without completing cytokinesis. This resulted in a gradual increase in the percentage of multinucleated cells in the culture. Time-lapse imaging of synchronized cultures revealed that control and Survivin-depleted cells arrested in mitosis in the presence of nocodazole; however, the latter failed to arrest in mitosis when treated with taxol. Immunofluorescence studies revealed that Survivindepleted cells were unable to stably maintain BubR1 at the kinetochores in the presence of either taxol or nocodazole. Our data reveal that Survivin is not required for the spindle assembly checkpoint when it is activated by the loss of microtubules. However, Survivin is required for the maintenance of the checkpoint when it is activated by taxol, which is generally thought to cause a loss of spindle tension.
History
Publication status
- Published
Journal
Journal of Cell ScienceISSN
0021-9533Publisher
Company of BiologistsExternal DOI
Issue
14Volume
116Page range
2987-2998Pages
12.0Department affiliated with
- Sussex Centre for Genome Damage Stability Publications
Notes
SPW directed the research and was corresponding author. This was one of the first papers to describe the consequence of knocking down survivin in human cells using siRNA, and to demonstrate that the chromosomal passenger proteins contribute to the stability of a robust spindle checkpoint.Full text available
- No
Peer reviewed?
- Yes
Legacy Posted Date
2012-02-06Usage metrics
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