Selective killing of transformed cells by cyclin/cyclin-dependent kinase 2 antagonists.

Recent studies identified a short peptide motif that serves as a docking site for cyclin/cyclin-dependent kinase (cdk) 2 complexes. Peptides containing this motif block the phosphorylation of substrates by cyclin A/cdk2 or cyclin E/cdk2. Here we report that cell membrane-permeable forms of such peptides preferentially induced transformed cells to undergo apoptosis relative to nontransformed cells. Deregulation of E2F family transcription factors is a common event during transformation and was sufficient to sensitize cells to the cyclin/cdk2 inhibitory peptides. These results suggest that deregulation of E2F and inhibition of cdk2 are synthetically lethal and provide a rationale for the development of cdk2 antagonists as antineoplastic agents.

[1]  A. Levine,et al.  p53 and E2F-1 cooperate to mediate apoptosis. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[2]  W. Kaelin,et al.  Role of the retinoblastoma protein in the pathogenesis of human cancer. , 1997, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[3]  Natalie A. Lissy,et al.  Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration , 1998, Nature Medicine.

[4]  L. Tsai,et al.  The cdk2 kinase is required for the G1-to-S transition in mammalian cells. , 1993, Oncogene.

[5]  B. Schulman,et al.  Substrate recruitment to cyclin-dependent kinase 2 by a multipurpose docking site on cyclin A. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[6]  C. Peng,et al.  Cyclin A/CDK2 binds directly to E2F-1 and inhibits the DNA-binding activity of E2F-1/DP-1 by phosphorylation , 1994, Molecular and cellular biology.

[7]  J. Bartek,et al.  Unrestricted cell cycling and cancer , 1995, Nature Medicine.

[8]  J. Hornung,et al.  Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line , 1988, The Journal of cell biology.

[9]  W. Kaelin,et al.  Deregulated transcription factor E2F-1 expression leads to S-phase entry and p53-mediated apoptosis. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[10]  K. Helin,et al.  Induction of DNA synthesis and apoptosis are separable functions of E2F-1. , 1997, Genes & development.

[11]  M. Vidal,et al.  The retinoblastoma protein binds to a family of E2F transcription factors , 1993, Molecular and cellular biology.

[12]  W. Lee,et al.  Deregulated expression of E2F-1 induces S-phase entry and leads to apoptosis , 1994, Molecular and cellular biology.

[13]  B. Dynlacht,et al.  Differential regulation of E2F transactivation by cyclin/cdk2 complexes. , 1994, Genes & development.

[14]  J Barsoum,et al.  Tat-mediated delivery of heterologous proteins into cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[15]  J. Nevins,et al.  Identification of positively and negatively acting elements regulating expression of the E2F2 gene in response to cell growth signals , 1997, Molecular and cellular biology.

[16]  Xin Lu,et al.  E2F1-induced apoptosis requires DNA binding but not transactivation and is inhibited by the retinoblastoma protein through direct interaction. , 1997, Genes & development.

[17]  J. Nevins,et al.  Expression of transcription factor E2F1 induces quiescent cells to enter S phase , 1993, Nature.

[18]  Philip D. Jeffrey,et al.  Crystal structure of the p27Kip1 cyclin-dependent-kinase inibitor bound to the cyclin A–Cdk2 complex , 1996, Nature.

[19]  W. Yung,et al.  Overexpression of E2F‐1 in glioma triggers apoptosis and suppresses tumor growth in vitro and in vivo , 1998, Nature Medicine.

[20]  D. Livingston,et al.  Cyclin A-kinase regulation of E2F-1 DNA binding function underlies suppression of an S phase checkpoint , 1995, Cell.

[21]  R Pepperkok,et al.  Regulation of the cell cycle by the cdk2 protein kinase in cultured human fibroblasts , 1993, The Journal of cell biology.

[22]  W. Sellers,et al.  Identification of a cyclin-cdk2 recognition motif present in substrates and p21-like cyclin-dependent kinase inhibitors , 1996, Molecular and cellular biology.

[23]  T. Nilsen,et al.  RNA-RNA interactions in the spliceosome: Unraveling the ties that bind , 1994, Cell.

[24]  Anindya Dutta,et al.  Cyclin-binding motifs are essential for the function of p21CIP1 , 1996, Molecular and cellular biology.

[25]  P. Farnham,et al.  Multiple DNA elements are required for the growth regulation of the mouse E2F1 promoter. , 1994, Genes & development.

[26]  J. Nevins,et al.  Autoregulatory control of E2F1 expression in response to positive and negative regulators of cell cycle progression. , 1994, Genes & development.

[27]  R. Fåhraeus,et al.  Inhibition of pRb phosphorylation and cell-cycle progression by a 20-residue peptide from p16CDKN2/INK4A , 1996, Current Biology.

[28]  C. Sherr.  Cancer Cell Cycles , 1996, Science.

[29]  W. Sellers,et al.  Transcription of the E2F-1 gene is rendered cell cycle dependent by E2F DNA-binding sites within its promoter , 1994, Molecular and cellular biology.