Mammalian S-phase checkpoint integrity is dependent on transformation status and purine deoxyribonucleosides

Stephen Downes, CZ Bachrati, SJ Devlin, M Tommasino, TJR Cutts, JV Watson, I Rasko, RT Johnson

    Research output: Contribution to journalArticle

    9 Citations (Scopus)

    Abstract

    In eukaryotic cells arrested in S-phase, checkpoint controls normally restrain mitosis until after replication. We have identified an array of previously unsuspected factors that modulate this restraint, using transformed hamster cells in which cycle controls are known to be altered in S-phase arrest. Arrested cells accumulate cyclin B, the regulatory partner of the mitotic p34(cdc2) kinase, which is normally not abundant until late G(2) phase; treatment of arrested cells with caffeine produces rapid S-phase condensation. We show here that such S-phase checkpoint slippage, as visualised through caffeine-dependent S-phase condensation, correlates with rodent origin and transformed status, is opposed by reverse transformation, and is favoured by c-src and opposed by wnt1 overexpression, Slippage is also dependent on a prolonged replicative arrest, and is favoured by arrest with hydroxyurea, which inhibits ribonucleotide reductase, This last is a key enzyme in deoxyribonucleotide synthesis, recently identified as a determinant of malignancy. Addition of deoxyribonucleosides shows that rapid S-phase condensation is suppressed by a novel checkpoint mechanism: purine (but not pyrimidine) deoxyribonucleosides, like reverse transformation, suppress cyclin B/p34(cdc2) activation by caffeine, but not cyclin B accumulation. Thus, ribonucleotide reductase has an unexpectedly complex role in mammalian cell cycle regulation: not only is it regulated in response to cycle progression, but its products can also reciprocally influence cell cycle control kinase activation.
    LanguageEnglish
    Pages1089-1096
    JournalJOURNAL OF CELL SCIENCE
    Volume113
    Issue number6
    Publication statusPublished - Mar 2000

    Fingerprint

    S Phase Cell Cycle Checkpoints
    Deoxyribonucleosides
    Cyclin B
    S Phase
    Caffeine
    Ribonucleotide Reductases
    Phosphotransferases
    Deoxyribonucleotides
    Hydroxyurea
    Eukaryotic Cells
    Cell Cycle Checkpoints
    Mitosis
    Cricetinae
    Rodentia
    Cell Cycle
    purine
    Enzymes
    Neoplasms

    Cite this

    Downes, S., Bachrati, CZ., Devlin, SJ., Tommasino, M., Cutts, TJR., Watson, JV., ... Johnson, RT. (2000). Mammalian S-phase checkpoint integrity is dependent on transformation status and purine deoxyribonucleosides. JOURNAL OF CELL SCIENCE, 113(6), 1089-1096.
    Downes, Stephen ; Bachrati, CZ ; Devlin, SJ ; Tommasino, M ; Cutts, TJR ; Watson, JV ; Rasko, I ; Johnson, RT. / Mammalian S-phase checkpoint integrity is dependent on transformation status and purine deoxyribonucleosides. In: JOURNAL OF CELL SCIENCE. 2000 ; Vol. 113, No. 6. pp. 1089-1096.
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    abstract = "In eukaryotic cells arrested in S-phase, checkpoint controls normally restrain mitosis until after replication. We have identified an array of previously unsuspected factors that modulate this restraint, using transformed hamster cells in which cycle controls are known to be altered in S-phase arrest. Arrested cells accumulate cyclin B, the regulatory partner of the mitotic p34(cdc2) kinase, which is normally not abundant until late G(2) phase; treatment of arrested cells with caffeine produces rapid S-phase condensation. We show here that such S-phase checkpoint slippage, as visualised through caffeine-dependent S-phase condensation, correlates with rodent origin and transformed status, is opposed by reverse transformation, and is favoured by c-src and opposed by wnt1 overexpression, Slippage is also dependent on a prolonged replicative arrest, and is favoured by arrest with hydroxyurea, which inhibits ribonucleotide reductase, This last is a key enzyme in deoxyribonucleotide synthesis, recently identified as a determinant of malignancy. Addition of deoxyribonucleosides shows that rapid S-phase condensation is suppressed by a novel checkpoint mechanism: purine (but not pyrimidine) deoxyribonucleosides, like reverse transformation, suppress cyclin B/p34(cdc2) activation by caffeine, but not cyclin B accumulation. Thus, ribonucleotide reductase has an unexpectedly complex role in mammalian cell cycle regulation: not only is it regulated in response to cycle progression, but its products can also reciprocally influence cell cycle control kinase activation.",
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    Downes, S, Bachrati, CZ, Devlin, SJ, Tommasino, M, Cutts, TJR, Watson, JV, Rasko, I & Johnson, RT 2000, 'Mammalian S-phase checkpoint integrity is dependent on transformation status and purine deoxyribonucleosides', JOURNAL OF CELL SCIENCE, vol. 113, no. 6, pp. 1089-1096.

    Mammalian S-phase checkpoint integrity is dependent on transformation status and purine deoxyribonucleosides. / Downes, Stephen; Bachrati, CZ; Devlin, SJ; Tommasino, M; Cutts, TJR; Watson, JV; Rasko, I; Johnson, RT.

    In: JOURNAL OF CELL SCIENCE, Vol. 113, No. 6, 03.2000, p. 1089-1096.

    Research output: Contribution to journalArticle

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    T1 - Mammalian S-phase checkpoint integrity is dependent on transformation status and purine deoxyribonucleosides

    AU - Downes, Stephen

    AU - Bachrati, CZ

    AU - Devlin, SJ

    AU - Tommasino, M

    AU - Cutts, TJR

    AU - Watson, JV

    AU - Rasko, I

    AU - Johnson, RT

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    N2 - In eukaryotic cells arrested in S-phase, checkpoint controls normally restrain mitosis until after replication. We have identified an array of previously unsuspected factors that modulate this restraint, using transformed hamster cells in which cycle controls are known to be altered in S-phase arrest. Arrested cells accumulate cyclin B, the regulatory partner of the mitotic p34(cdc2) kinase, which is normally not abundant until late G(2) phase; treatment of arrested cells with caffeine produces rapid S-phase condensation. We show here that such S-phase checkpoint slippage, as visualised through caffeine-dependent S-phase condensation, correlates with rodent origin and transformed status, is opposed by reverse transformation, and is favoured by c-src and opposed by wnt1 overexpression, Slippage is also dependent on a prolonged replicative arrest, and is favoured by arrest with hydroxyurea, which inhibits ribonucleotide reductase, This last is a key enzyme in deoxyribonucleotide synthesis, recently identified as a determinant of malignancy. Addition of deoxyribonucleosides shows that rapid S-phase condensation is suppressed by a novel checkpoint mechanism: purine (but not pyrimidine) deoxyribonucleosides, like reverse transformation, suppress cyclin B/p34(cdc2) activation by caffeine, but not cyclin B accumulation. Thus, ribonucleotide reductase has an unexpectedly complex role in mammalian cell cycle regulation: not only is it regulated in response to cycle progression, but its products can also reciprocally influence cell cycle control kinase activation.

    AB - In eukaryotic cells arrested in S-phase, checkpoint controls normally restrain mitosis until after replication. We have identified an array of previously unsuspected factors that modulate this restraint, using transformed hamster cells in which cycle controls are known to be altered in S-phase arrest. Arrested cells accumulate cyclin B, the regulatory partner of the mitotic p34(cdc2) kinase, which is normally not abundant until late G(2) phase; treatment of arrested cells with caffeine produces rapid S-phase condensation. We show here that such S-phase checkpoint slippage, as visualised through caffeine-dependent S-phase condensation, correlates with rodent origin and transformed status, is opposed by reverse transformation, and is favoured by c-src and opposed by wnt1 overexpression, Slippage is also dependent on a prolonged replicative arrest, and is favoured by arrest with hydroxyurea, which inhibits ribonucleotide reductase, This last is a key enzyme in deoxyribonucleotide synthesis, recently identified as a determinant of malignancy. Addition of deoxyribonucleosides shows that rapid S-phase condensation is suppressed by a novel checkpoint mechanism: purine (but not pyrimidine) deoxyribonucleosides, like reverse transformation, suppress cyclin B/p34(cdc2) activation by caffeine, but not cyclin B accumulation. Thus, ribonucleotide reductase has an unexpectedly complex role in mammalian cell cycle regulation: not only is it regulated in response to cycle progression, but its products can also reciprocally influence cell cycle control kinase activation.

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    Downes S, Bachrati CZ, Devlin SJ, Tommasino M, Cutts TJR, Watson JV et al. Mammalian S-phase checkpoint integrity is dependent on transformation status and purine deoxyribonucleosides. JOURNAL OF CELL SCIENCE. 2000 Mar;113(6):1089-1096.