Coordinate modulation of Sp1, NF-kappa B, and p53 in confluent human malignant melanoma cells after ionizing radiation

Chin Rang Yang, Carmell Wilson Van Patten, Sarah M. Planchon, Shelly M. Wuerzberger-Davis, Thomas W. Davis, Scott Cuthill, Shigeki Miyamoto, David A. Boothman

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

Regulation of transcriptional responses in growth-arrested human cells under conditions that promote potentially lethal damage repair after ionizing radiation (IR) is poorly understood. Sp1/retinoblastoma control protein (RCP) DNA binding increased within 30 min and peaked at 2-4 h after IR (450-600 cGy) in confluent radioresistant human malignant melanoma (U1-Mel) cells. Increased phosphorylation of Sp1 directly corresponded to Sp1/RCP binding and immediate-early gene induction, whereas pRb remained hypophosphorylated. Transfection of U1-Mel cells with the human papillomavirus E7 gene abrogated Sp1/RCP induction and G0/G1 cell cycle checkpoint arrest responses, increased apoptosis and radiosensitivity, and augmented genetic instability (i.e., increased polyploidy cells) after IR. Increased NF-κB DNA binding in U1-Mel cells after IR treatment lasted much longer (i.e., >20 h). U1-Mel cells overexpressing dominant-negative IκBα S32/36A mutant protein were significantly more resistant to IR exposure and retained both G2/M and G0/G1 cell cycle checkpoint responses without significant genetic instability (i.e., polyploid cell populations were not observed). Nuclear p53 protein levels and DNA binding activity increased only after high doses of IR (> 1200 cGy). Disruption of p53 responses in U1-Mel cells by E6 transfection also abrogated G0/G1 cell cycle checkpoint arrest responses and increased polyploidy after IR, but did not alter radiosensitivity. These data suggest that abrogation of individual components of this coordinate IR-activated transcription factor response may lead to divergent alterations in cell cycle checkpoints, genomic instability, apoptosis, and survival. Such coordinate transcription factor activation in human cancer cells is reminiscent of prokaryotic SOS responses, and further elucidation of these events should shed light on the initial molecular events in the chromosome instability phenotype.

Original languageEnglish (US)
Pages (from-to)379-390
Number of pages12
JournalFASEB Journal
Volume14
Issue number2
StatePublished - 2000

Fingerprint

NF-kappa B
Ionizing radiation
melanoma
Ionizing Radiation
ionizing radiation
Melanoma
G1 Phase Cell Cycle Checkpoints
Modulation
Cells
Polyploidy
polyploidy
cells
Retinoblastoma Protein
Radiation Tolerance
transfection
Transfection
Retinoblastoma Binding Proteins
DNA
Transcription Factors
apoptosis

Keywords

  • Aneuploid/polyploid
  • Apoptosis
  • Genomic instability
  • Ionizing radiation
  • NF-κB
  • P53
  • PLDR
  • Retinoblastoma control proteins
  • Spl

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Yang, C. R., Patten, C. W. V., Planchon, S. M., Wuerzberger-Davis, S. M., Davis, T. W., Cuthill, S., ... Boothman, D. A. (2000). Coordinate modulation of Sp1, NF-kappa B, and p53 in confluent human malignant melanoma cells after ionizing radiation. FASEB Journal, 14(2), 379-390.

Coordinate modulation of Sp1, NF-kappa B, and p53 in confluent human malignant melanoma cells after ionizing radiation. / Yang, Chin Rang; Patten, Carmell Wilson Van; Planchon, Sarah M.; Wuerzberger-Davis, Shelly M.; Davis, Thomas W.; Cuthill, Scott; Miyamoto, Shigeki; Boothman, David A.

In: FASEB Journal, Vol. 14, No. 2, 2000, p. 379-390.

Research output: Contribution to journalArticle

Yang, CR, Patten, CWV, Planchon, SM, Wuerzberger-Davis, SM, Davis, TW, Cuthill, S, Miyamoto, S & Boothman, DA 2000, 'Coordinate modulation of Sp1, NF-kappa B, and p53 in confluent human malignant melanoma cells after ionizing radiation', FASEB Journal, vol. 14, no. 2, pp. 379-390.
Yang CR, Patten CWV, Planchon SM, Wuerzberger-Davis SM, Davis TW, Cuthill S et al. Coordinate modulation of Sp1, NF-kappa B, and p53 in confluent human malignant melanoma cells after ionizing radiation. FASEB Journal. 2000;14(2):379-390.
Yang, Chin Rang ; Patten, Carmell Wilson Van ; Planchon, Sarah M. ; Wuerzberger-Davis, Shelly M. ; Davis, Thomas W. ; Cuthill, Scott ; Miyamoto, Shigeki ; Boothman, David A. / Coordinate modulation of Sp1, NF-kappa B, and p53 in confluent human malignant melanoma cells after ionizing radiation. In: FASEB Journal. 2000 ; Vol. 14, No. 2. pp. 379-390.
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AU - Wuerzberger-Davis, Shelly M.

AU - Davis, Thomas W.

AU - Cuthill, Scott

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