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Caspase activity is not required for the mitotic checkpoint or mitotic slippage in human cells
Biochemical studies suggest that caspase activity is required for a functional mitotic checkpoint (MC) and mitotic slippage. To test this directly, we followed nontransformed human telomerase immortalized human retinal pigment epithelia (RPE-1) cells through mitosis after inhibiting or depleting se...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2011
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3135473/ https://www.ncbi.nlm.nih.gov/pubmed/21613548 http://dx.doi.org/10.1091/mbc.E11-03-0228 |
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author | Lee, Kyunghee Kenny, Alison E. Rieder, Conly L. |
author_facet | Lee, Kyunghee Kenny, Alison E. Rieder, Conly L. |
author_sort | Lee, Kyunghee |
collection | PubMed |
description | Biochemical studies suggest that caspase activity is required for a functional mitotic checkpoint (MC) and mitotic slippage. To test this directly, we followed nontransformed human telomerase immortalized human retinal pigment epithelia (RPE-1) cells through mitosis after inhibiting or depleting selected caspases. We found that inhibiting caspases individually, in combination, or in toto did not affect the duration or fidelity of mitosis in otherwise untreated cells. When satisfaction of the MC was prevented with 500 nM nocodazole or 2.5 μM dimethylenastron (an Eg5 inhibitor), 92–100% of RPE-1 cells slipped from mitosis in the presence of pan-caspase inhibitors or after simultaneously depleting caspase-3 and -9, and they did so with the same kinetics (∼21–22 h) as after treatment with nocodazole or Eg5 inhibitors alone. Surprisingly, inhibiting or depleting caspase-9 alone doubled the number of nocodazole-treated, but not Eg5-inhibited, cells that died in mitosis. In addition, inhibiting or depleting caspase-9 and -3 together accelerated the rate of slippage ∼40% (to ∼13–15 h). Finally, nocodazole-treated cells that recently slipped through mitosis in the presence or absence of pan-caspase inhibitors contained numerous BubR1 foci in their nuclei. From these data, we conclude that caspase activity is not required for a functional MC or for mitotic slippage. |
format | Online Article Text |
id | pubmed-3135473 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-31354732011-09-30 Caspase activity is not required for the mitotic checkpoint or mitotic slippage in human cells Lee, Kyunghee Kenny, Alison E. Rieder, Conly L. Mol Biol Cell Articles Biochemical studies suggest that caspase activity is required for a functional mitotic checkpoint (MC) and mitotic slippage. To test this directly, we followed nontransformed human telomerase immortalized human retinal pigment epithelia (RPE-1) cells through mitosis after inhibiting or depleting selected caspases. We found that inhibiting caspases individually, in combination, or in toto did not affect the duration or fidelity of mitosis in otherwise untreated cells. When satisfaction of the MC was prevented with 500 nM nocodazole or 2.5 μM dimethylenastron (an Eg5 inhibitor), 92–100% of RPE-1 cells slipped from mitosis in the presence of pan-caspase inhibitors or after simultaneously depleting caspase-3 and -9, and they did so with the same kinetics (∼21–22 h) as after treatment with nocodazole or Eg5 inhibitors alone. Surprisingly, inhibiting or depleting caspase-9 alone doubled the number of nocodazole-treated, but not Eg5-inhibited, cells that died in mitosis. In addition, inhibiting or depleting caspase-9 and -3 together accelerated the rate of slippage ∼40% (to ∼13–15 h). Finally, nocodazole-treated cells that recently slipped through mitosis in the presence or absence of pan-caspase inhibitors contained numerous BubR1 foci in their nuclei. From these data, we conclude that caspase activity is not required for a functional MC or for mitotic slippage. The American Society for Cell Biology 2011-07-15 /pmc/articles/PMC3135473/ /pubmed/21613548 http://dx.doi.org/10.1091/mbc.E11-03-0228 Text en © 2011 Lee et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology. |
spellingShingle | Articles Lee, Kyunghee Kenny, Alison E. Rieder, Conly L. Caspase activity is not required for the mitotic checkpoint or mitotic slippage in human cells |
title | Caspase activity is not required for the mitotic checkpoint or mitotic slippage in human cells |
title_full | Caspase activity is not required for the mitotic checkpoint or mitotic slippage in human cells |
title_fullStr | Caspase activity is not required for the mitotic checkpoint or mitotic slippage in human cells |
title_full_unstemmed | Caspase activity is not required for the mitotic checkpoint or mitotic slippage in human cells |
title_short | Caspase activity is not required for the mitotic checkpoint or mitotic slippage in human cells |
title_sort | caspase activity is not required for the mitotic checkpoint or mitotic slippage in human cells |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3135473/ https://www.ncbi.nlm.nih.gov/pubmed/21613548 http://dx.doi.org/10.1091/mbc.E11-03-0228 |
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