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PMID: 18025303 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Mitosis persists in the absence of Cdk1 activity when proteolysis or protein phosphatase activity is suppressed.

The Journal of cell biology ·Vol. 179 ·No. 4 ·2007-11-19 ·Pages 671-85

Skoufias DA, Indorato RL, Lacroix F, Panopoulos A, Margolis RL

Abstract

Cellular transition to anaphase and mitotic exit has been linked to the loss of cyclin-dependent kinase 1 (Cdk1) kinase activity as a result of anaphase-promoting complex/cyclosome (APC/C)-dependent specific degradation of its cyclin B1 subunit. Cdk1 inhibition by roscovitine is known to induce premature mitotic exit, whereas inhibition of the APC/C-dependent degradation of cyclin B1 by MG132 induces mitotic arrest. In this study, we find that combining both drugs causes prolonged mitotic arrest in the absence of Cdk1 activity. Different Cdk1 and proteasome inhibitors produce similar results, indicating that the effect is not drug specific. We verify mitotic status by the retention of mitosis-specific markers and Cdk1 phosphorylation substrates, although cells can undergo late mitotic furrowing while still in mitosis. Overall, we conclude that continuous Cdk1 activity is not essential to maintain the mitotic state and that phosphatase activity directed at Cdk1 substrates is largely quiescent during mitosis. Furthermore, the degradation of a protein other than cyclin B1 is essential to activate a phosphatase that, in turn, enables mitotic exit.

MeSH Terms
2-Aminopurine/analogs & derivatives,pharmacology CDC2 Protein Kinase/antagonists & inhibitors Coloring Agents Cysteine Proteinase Inhibitors/pharmacology Drug Interactions Enzyme Inhibitors/pharmacology Fluorescein-5-isothiocyanate Fluorescent Antibody Technique, Indirect Fluorescent Dyes HCT116 Cells HeLa Cells Humans Hydrolysis Lactams/pharmacology Leupeptins/pharmacology Mitosis/drug effects Phosphoprotein Phosphatases/antagonists & inhibitors Propidium Protein Kinase Inhibitors/pharmacology Purines/pharmacology Roscovitine
Chemicals
6-((3-chloro)anilino)-2-(isopropyl-2-hydroxyethylamino)-9-isopropylpurine AM 114 Coloring Agents Cysteine Proteinase Inhibitors Enzyme Inhibitors Fluorescent Dyes Lactams Leupeptins N(2)-(2-aminocyclohexyl)-N(6)-(3-chlorophenyl)-9-ethyl-9H-purine-2,6-diamine Protein Kinase Inhibitors Purines Roscovitine Propidium 2-Aminopurine CDC2 Protein Kinase Phosphoprotein Phosphatases Fluorescein-5-isothiocyanate benzyloxycarbonylleucyl-leucyl-leucine aldehyde
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Skoufias Dimitrios A
Institut de Biologie Structurale Jean-Pierre Ebel, Atomic Energy Commission/Centre National de la Recherche Scientifique, 38027 Grenoble, Cedex 1, France. dimitrios.skoufias@ibs.fr
Indorato Rose-Laure
Lacroix Françoise
Panopoulos Andreas
Margolis Robert L
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.200704117
Email
dimitrios.skoufias@ibs.fr
Published
2007-11-19
Pages
671-85
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2080905
Subset
IM
Grants
NIGMS NIH HHS · R01 GM068107 · United States
NIGMS NIH HHS · GM068107 · United States
External Links
PubMed source DOI: 10.1083/jcb.200704117 PMC Full Text View journal details
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