Induction of p21(WAF1/CIP1) and inhibition of Cdk2 mediated by the tumor suppressor p16(INK4α)

Jayashree Mitra, Charlotte Y. Dai, Kumaravel Somasundaram, Wafik S. El-Deiry, Kapaettu Satyamoorthy, Meenhard Herlyn, Greg H. Enders

    Research output: Contribution to journalArticle

    106 Citations (Scopus)

    Abstract

    The tumor suppressor p16(INK4α) inhibits cyclin-dependent kinases 4 and 6. This activates the retinoblastoma protein (pRB) and, through incompletely understood events, arrests the cell division cycle. To permit biochemical analysis of the arrest, we generated U2-OS osteogenic sarcoma cell clones in which p16 transcription could be induced. In these clones, binding of p16 to cdk4 and cdk6 abrogated binding of cyclin D1, p27(KIP1), and p21(WAF1/CIP1). Concomitantly, the total cellular level of p21 increased severalfold via a posttranscriptional mechanism. Most cyclin E-cdk2 complexes associated with p21 and became inactive, expression of cyclin A was curtailed, and DNA synthesis was strongly inhibited. Induction of p21 alone, in a sibling clone, to the level observed during p16 induction substantially reproduced these effects. Overexpression of either cyclin E or A prevented p16 from mediating arrest. We then extended these studies to HCT 116 colorectal carcinoma cells and a p21-null clone derived by homologous recombination. In the parental cells, p16 expression also augmented total cellular and cdk2-bound p21. Moreover, p16 strongly inhibited DNA synthesis in the parental cells but not in the p21-null derivative. These findings indicate that p21-mediated inhibition of cdk2 contributes to the cell cycle arrest imposed by p16 and is a potential point of cooperation between the p16/pRB and p14(ARF)/p53 tumor suppressor pathways.

    Original languageEnglish
    Pages (from-to)3916-3928
    Number of pages13
    JournalMolecular and Cellular Biology
    Volume19
    Issue number5
    Publication statusPublished - 05-1999

    Fingerprint

    Clone Cells
    Cyclin A
    Cyclin E
    Neoplasms
    Cyclin-Dependent Kinase 6
    Tumor Suppressor Protein p14ARF
    Cyclin-Dependent Kinase 4
    Retinoblastoma Protein
    Homologous Recombination
    DNA
    Cyclin D1
    Osteosarcoma
    Cell Cycle Checkpoints
    Colorectal Neoplasms
    Cell Cycle

    All Science Journal Classification (ASJC) codes

    • Molecular Biology
    • Genetics
    • Cell Biology

    Cite this

    Mitra, J., Dai, C. Y., Somasundaram, K., El-Deiry, W. S., Satyamoorthy, K., Herlyn, M., & Enders, G. H. (1999). Induction of p21(WAF1/CIP1) and inhibition of Cdk2 mediated by the tumor suppressor p16(INK4α). Molecular and Cellular Biology, 19(5), 3916-3928.
    Mitra, Jayashree ; Dai, Charlotte Y. ; Somasundaram, Kumaravel ; El-Deiry, Wafik S. ; Satyamoorthy, Kapaettu ; Herlyn, Meenhard ; Enders, Greg H. / Induction of p21(WAF1/CIP1) and inhibition of Cdk2 mediated by the tumor suppressor p16(INK4α). In: Molecular and Cellular Biology. 1999 ; Vol. 19, No. 5. pp. 3916-3928.
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    abstract = "The tumor suppressor p16(INK4α) inhibits cyclin-dependent kinases 4 and 6. This activates the retinoblastoma protein (pRB) and, through incompletely understood events, arrests the cell division cycle. To permit biochemical analysis of the arrest, we generated U2-OS osteogenic sarcoma cell clones in which p16 transcription could be induced. In these clones, binding of p16 to cdk4 and cdk6 abrogated binding of cyclin D1, p27(KIP1), and p21(WAF1/CIP1). Concomitantly, the total cellular level of p21 increased severalfold via a posttranscriptional mechanism. Most cyclin E-cdk2 complexes associated with p21 and became inactive, expression of cyclin A was curtailed, and DNA synthesis was strongly inhibited. Induction of p21 alone, in a sibling clone, to the level observed during p16 induction substantially reproduced these effects. Overexpression of either cyclin E or A prevented p16 from mediating arrest. We then extended these studies to HCT 116 colorectal carcinoma cells and a p21-null clone derived by homologous recombination. In the parental cells, p16 expression also augmented total cellular and cdk2-bound p21. Moreover, p16 strongly inhibited DNA synthesis in the parental cells but not in the p21-null derivative. These findings indicate that p21-mediated inhibition of cdk2 contributes to the cell cycle arrest imposed by p16 and is a potential point of cooperation between the p16/pRB and p14(ARF)/p53 tumor suppressor pathways.",
    author = "Jayashree Mitra and Dai, {Charlotte Y.} and Kumaravel Somasundaram and El-Deiry, {Wafik S.} and Kapaettu Satyamoorthy and Meenhard Herlyn and Enders, {Greg H.}",
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    Mitra, J, Dai, CY, Somasundaram, K, El-Deiry, WS, Satyamoorthy, K, Herlyn, M & Enders, GH 1999, 'Induction of p21(WAF1/CIP1) and inhibition of Cdk2 mediated by the tumor suppressor p16(INK4α)', Molecular and Cellular Biology, vol. 19, no. 5, pp. 3916-3928.

    Induction of p21(WAF1/CIP1) and inhibition of Cdk2 mediated by the tumor suppressor p16(INK4α). / Mitra, Jayashree; Dai, Charlotte Y.; Somasundaram, Kumaravel; El-Deiry, Wafik S.; Satyamoorthy, Kapaettu; Herlyn, Meenhard; Enders, Greg H.

    In: Molecular and Cellular Biology, Vol. 19, No. 5, 05.1999, p. 3916-3928.

    Research output: Contribution to journalArticle

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    AU - Mitra, Jayashree

    AU - Dai, Charlotte Y.

    AU - Somasundaram, Kumaravel

    AU - El-Deiry, Wafik S.

    AU - Satyamoorthy, Kapaettu

    AU - Herlyn, Meenhard

    AU - Enders, Greg H.

    PY - 1999/5

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