Partial inhibition of Cdk1 in G2 phase overrides the SAC and decouples mitotic events
Identifieur interne : 001861 ( Ncbi/Curation ); précédent : 001860; suivant : 001862Partial inhibition of Cdk1 in G2 phase overrides the SAC and decouples mitotic events
Auteurs : Rachael A. Mccloy [Australie] ; Samuel Rogers [Australie] ; C Elizabeth Caldon [Australie] ; Thierry Lorca [France] ; Anna Castro [France] ; Andrew Burgess [Australie]Source :
- Cell Cycle [ 1538-4101 ] ; 2014.
Descripteurs français
- KwdFr :
- Acide okadaïque (pharmacologie), Appareil du fuseau (physiologie), Appareil du fuseau (ultrastructure), Cellules HeLa, Centrosome (), Centrosome (métabolisme), Cytocinèse, Humains, Mitose, Phase G2, Phosphorylation, Polyploïdie, Protéine-kinase CDC2 (antagonistes et inhibiteurs), Protéine-kinase CDC2 (métabolisme), Quinoléines (pharmacologie), Thiazoles (pharmacologie).
- MESH :
- antagonistes et inhibiteurs : Protéine-kinase CDC2.
- métabolisme : Centrosome, Protéine-kinase CDC2.
- pharmacologie : Acide okadaïque, Quinoléines, Thiazoles.
- physiologie : Appareil du fuseau.
- Appareil du fuseau, Cellules HeLa, Centrosome, Cytocinèse, Humains, Mitose, Phase G2, Phosphorylation, Polyploïdie.
English descriptors
- KwdEn :
- CDC2 Protein Kinase (antagonists & inhibitors), CDC2 Protein Kinase (metabolism), Centrosome (drug effects), Centrosome (metabolism), Cytokinesis, G2 Phase, HeLa Cells, Humans, Mitosis, Okadaic Acid (pharmacology), Phosphorylation, Polyploidy, Quinolines (pharmacology), Spindle Apparatus (physiology), Spindle Apparatus (ultrastructure), Thiazoles (pharmacology).
- MESH :
- chemical , antagonists & inhibitors : CDC2 Protein Kinase.
- chemical , metabolism : CDC2 Protein Kinase.
- drug effects : Centrosome.
- metabolism : Centrosome.
- chemical , pharmacology : Okadaic Acid, Quinolines, Thiazoles.
- physiology : Spindle Apparatus.
- ultrastructure : Spindle Apparatus.
- Cytokinesis, G2 Phase, HeLa Cells, Humans, Mitosis, Phosphorylation, Polyploidy.
Abstract
Entry and progression through mitosis has traditionally been linked directly to the activity of cyclin-dependent kinase 1 (Cdk1). In this study we utilized low doses of the Cdk1-specific inhibitor, RO3306 from early G2 phase onwards. Addition of low doses of RO3306 in G2 phase induced minor chromosome congression and segregation defects. In contrast, mild doses of RO3306 during G2 phase resulted in cells entering an aberrant mitosis, with cells fragmenting centrosomes and failing to fully disassemble the nuclear envelope. Cells often underwent cytokinesis and metaphase simultaneously, despite the presence of an active spindle assembly checkpoint, which prevented degradation of cyclin B1 and securin, resulting in the random partitioning of whole chromosomes. This highly aberrant mitosis produced a significant increase in the proportion of viable polyploid cells present up to 3 days post-treatment. Furthermore, cells treated with medium doses of RO3306 were only able to reach the threshold of Cdk1 substrate phosphorylation required to initiate nuclear envelope breakdown, but failed to reach the levels of phosphorylation required to correctly complete pro-metaphase. Treatment with low doses of Okadaic acid, which primarily inhibits PP2A, rescued the mitotic defects and increased the number of cells that completed a normal mitosis. This supports the current model that PP2A is the primary phosphatase that counterbalances the activity of Cdk1 during mitosis. Taken together these results show that continuous and subtle disruption of Cdk1 activity from G2 phase onwards has deleterious consequences on mitotic progression by disrupting the balance between Cdk1 and PP2A.
Url:
DOI: 10.4161/cc.28401
PubMed: 24626186
PubMed Central: 4050138
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PMC:4050138Le document en format XML
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Mccloy</name><affiliation wicri:level="1"><nlm:aff id="A1">The Kinghorn Cancer Centre; Garvan Institute of Medical Research; Sydney, New South Wales, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>The Kinghorn Cancer Centre; Garvan Institute of Medical Research; Sydney, New South Wales</wicri:regionArea><wicri:noRegion>New South Wales</wicri:noRegion></affiliation></author><author><name sortKey="Rogers, Samuel" sort="Rogers, Samuel" uniqKey="Rogers S" first="Samuel" last="Rogers">Samuel Rogers</name><affiliation wicri:level="1"><nlm:aff id="A1">The Kinghorn Cancer Centre; Garvan Institute of Medical Research; Sydney, New South Wales, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>The Kinghorn Cancer Centre; Garvan Institute of Medical Research; Sydney, New South Wales</wicri:regionArea><wicri:noRegion>New South Wales</wicri:noRegion></affiliation></author><author><name sortKey="Caldon, C Elizabeth" sort="Caldon, C Elizabeth" uniqKey="Caldon C" first="C Elizabeth" last="Caldon">C Elizabeth Caldon</name><affiliation wicri:level="1"><nlm:aff id="A1">The Kinghorn Cancer Centre; Garvan Institute of Medical Research; Sydney, New South Wales, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>The Kinghorn Cancer Centre; Garvan Institute of Medical Research; Sydney, New South Wales</wicri:regionArea><wicri:noRegion>New South Wales</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="A2">St. Vincent’s Clinical School; Faculty of Medicine; UNSW; Sydney, New South Wales, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>St. Vincent’s Clinical School; Faculty of Medicine; UNSW; Sydney, New South Wales</wicri:regionArea><wicri:noRegion>New South Wales</wicri:noRegion></affiliation></author><author><name sortKey="Lorca, Thierry" sort="Lorca, Thierry" uniqKey="Lorca T" first="Thierry" last="Lorca">Thierry Lorca</name><affiliation wicri:level="1"><nlm:aff id="A3">Universités Montpellier 2 et 1; Centre de Recherche de Biochimie Macromoléculaire; CNRS UMR 5237; Montpellier, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Universités Montpellier 2 et 1; Centre de Recherche de Biochimie Macromoléculaire; CNRS UMR 5237; Montpellier</wicri:regionArea></affiliation></author><author><name sortKey="Castro, Anna" sort="Castro, Anna" uniqKey="Castro A" first="Anna" last="Castro">Anna Castro</name><affiliation wicri:level="1"><nlm:aff id="A3">Universités Montpellier 2 et 1; Centre de Recherche de Biochimie Macromoléculaire; CNRS UMR 5237; Montpellier, France</nlm:aff><country xml:lang="fr">France</country><wicri:regionArea>Universités Montpellier 2 et 1; Centre de Recherche de Biochimie Macromoléculaire; CNRS UMR 5237; Montpellier</wicri:regionArea></affiliation></author><author><name sortKey="Burgess, Andrew" sort="Burgess, Andrew" uniqKey="Burgess A" first="Andrew" last="Burgess">Andrew Burgess</name><affiliation wicri:level="1"><nlm:aff id="A1">The Kinghorn Cancer Centre; Garvan Institute of Medical Research; Sydney, New South Wales, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>The Kinghorn Cancer Centre; Garvan Institute of Medical Research; Sydney, New South Wales</wicri:regionArea><wicri:noRegion>New South Wales</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="A2">St. Vincent’s Clinical School; Faculty of Medicine; UNSW; Sydney, New South Wales, Australia</nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>St. Vincent’s Clinical School; Faculty of Medicine; UNSW; Sydney, New South Wales</wicri:regionArea><wicri:noRegion>New South Wales</wicri:noRegion></affiliation></author></analytic><series><title level="j">Cell Cycle</title><idno type="ISSN">1538-4101</idno><idno type="eISSN">1551-4005</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>CDC2 Protein Kinase (antagonists & inhibitors)</term><term>CDC2 Protein Kinase (metabolism)</term><term>Centrosome (drug effects)</term><term>Centrosome (metabolism)</term><term>Cytokinesis</term><term>G2 Phase</term><term>HeLa Cells</term><term>Humans</term><term>Mitosis</term><term>Okadaic Acid (pharmacology)</term><term>Phosphorylation</term><term>Polyploidy</term><term>Quinolines (pharmacology)</term><term>Spindle Apparatus (physiology)</term><term>Spindle Apparatus (ultrastructure)</term><term>Thiazoles (pharmacology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Acide okadaïque (pharmacologie)</term><term>Appareil du fuseau (physiologie)</term><term>Appareil du fuseau (ultrastructure)</term><term>Cellules HeLa</term><term>Centrosome ()</term><term>Centrosome (métabolisme)</term><term>Cytocinèse</term><term>Humains</term><term>Mitose</term><term>Phase G2</term><term>Phosphorylation</term><term>Polyploïdie</term><term>Protéine-kinase CDC2 (antagonistes et inhibiteurs)</term><term>Protéine-kinase CDC2 (métabolisme)</term><term>Quinoléines (pharmacologie)</term><term>Thiazoles (pharmacologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="antagonists & inhibitors" xml:lang="en"><term>CDC2 Protein Kinase</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>CDC2 Protein Kinase</term></keywords><keywords scheme="MESH" qualifier="antagonistes et inhibiteurs" xml:lang="fr"><term>Protéine-kinase CDC2</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Centrosome</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Centrosome</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Centrosome</term><term>Protéine-kinase CDC2</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Acide okadaïque</term><term>Quinoléines</term><term>Thiazoles</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Okadaic Acid</term><term>Quinolines</term><term>Thiazoles</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Appareil du fuseau</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Spindle Apparatus</term></keywords><keywords scheme="MESH" qualifier="ultrastructure" xml:lang="en"><term>Spindle Apparatus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cytokinesis</term><term>G2 Phase</term><term>HeLa Cells</term><term>Humans</term><term>Mitosis</term><term>Phosphorylation</term><term>Polyploidy</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Appareil du fuseau</term><term>Cellules HeLa</term><term>Centrosome</term><term>Cytocinèse</term><term>Humains</term><term>Mitose</term><term>Phase G2</term><term>Phosphorylation</term><term>Polyploïdie</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Entry and progression through mitosis has traditionally been linked directly to the activity of cyclin-dependent kinase 1 (Cdk1). In this study we utilized low doses of the Cdk1-specific inhibitor, RO3306 from early G<sub>2</sub> phase onwards. Addition of low doses of RO3306 in G<sub>2</sub> phase induced minor chromosome congression and segregation defects. In contrast, mild doses of RO3306 during G<sub>2</sub> phase resulted in cells entering an aberrant mitosis, with cells fragmenting centrosomes and failing to fully disassemble the nuclear envelope. Cells often underwent cytokinesis and metaphase simultaneously, despite the presence of an active spindle assembly checkpoint, which prevented degradation of cyclin B1 and securin, resulting in the random partitioning of whole chromosomes. This highly aberrant mitosis produced a significant increase in the proportion of viable polyploid cells present up to 3 days post-treatment. Furthermore, cells treated with medium doses of RO3306 were only able to reach the threshold of Cdk1 substrate phosphorylation required to initiate nuclear envelope breakdown, but failed to reach the levels of phosphorylation required to correctly complete pro-metaphase. Treatment with low doses of Okadaic acid, which primarily inhibits PP2A, rescued the mitotic defects and increased the number of cells that completed a normal mitosis. This supports the current model that PP2A is the primary phosphatase that counterbalances the activity of Cdk1 during mitosis. Taken together these results show that continuous and subtle disruption of Cdk1 activity from G<sub>2</sub> phase onwards has deleterious consequences on mitotic progression by disrupting the balance between Cdk1 and PP2A.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Sherr, Cj" uniqKey="Sherr C">CJ Sherr</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Satyanarayana, A" uniqKey="Satyanarayana A">A Satyanarayana</name></author><author><name sortKey="Kaldis, P" uniqKey="Kaldis P">P Kaldis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pines, J" uniqKey="Pines J">J Pines</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Holt, Lj" uniqKey="Holt L">LJ Holt</name></author><author><name sortKey="Tuch, Bb" uniqKey="Tuch B">BB Tuch</name></author><author><name sortKey="Villen, J" uniqKey="Villen J">J 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Gan</name></author><author><name sortKey="Sanz Flores, M" uniqKey="Sanz Flores M">M Sanz-Flores</name></author><author><name sortKey="Trakala, M" uniqKey="Trakala M">M Trakala</name></author><author><name sortKey="Ruiz Torres, M" uniqKey="Ruiz Torres M">M Ruiz-Torres</name></author><author><name sortKey="Lorca, T" uniqKey="Lorca T">T Lorca</name></author><author><name sortKey="Castro, A" uniqKey="Castro A">A Castro</name></author><author><name sortKey="Malumbres, M" uniqKey="Malumbres M">M Malumbres</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Voets, E" uniqKey="Voets E">E Voets</name></author><author><name sortKey="Wolthuis, Rmf" uniqKey="Wolthuis R">RMF Wolthuis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gabrielli, Bg" uniqKey="Gabrielli B">BG Gabrielli</name></author><author><name sortKey="De Souza, Cp" uniqKey="De Souza C">CP De Souza</name></author><author><name sortKey="Tonks, Id" uniqKey="Tonks I">ID Tonks</name></author><author><name sortKey="Clark, Jm" uniqKey="Clark J">JM Clark</name></author><author><name sortKey="Hayward, Nk" uniqKey="Hayward N">NK Hayward</name></author><author><name sortKey="Ellem, Ka" uniqKey="Ellem K">KA Ellem</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Burgess, A" uniqKey="Burgess A">A Burgess</name></author><author><name sortKey="Lorca, T" uniqKey="Lorca T">T Lorca</name></author><author><name sortKey="Castro, A" uniqKey="Castro A">A Castro</name></author></analytic></biblStruct></listBibl></div1></back></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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