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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...

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Autores principales: Lee, Kyunghee, Kenny, Alison E., Rieder, Conly L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2011
Materias:
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.
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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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