Functional analysis of p53 binding under differential stresses

Adam Krieg, Ester M. Hammond, Amato J. Giaccia

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Hypoxia and DNA damage stabilize the p53 protein, but the subsequent effect that each stress has on transcriptional regulation of known p53 target genes is variable. We have used chromatin immunoprecipitation followed by CpG island (CGI) microarray hybridization to identify promoters bound by p53 under both DNA-damaging and non-DNA-damaging conditions in HCT116 cells. Using gene-specific PCR analysis, we have verified an association with CGIs of the highest enrichment (>2.5-fold) (REV3L, XPMC2H, HNRPUL1, TOR1AIP1, glutathione peroxidase 1, and SCFD2), with CGIs of intermediate enrichment (<2.2-fold) (COX7A2L, SYVN1, and JAG2), and with CGIs of low enrichment (>2.0-fold) (MYC and PCNA). We found little difference in promoter binding when p53 is stabilized by these two distinctly different stresses. However, expression of these genes varies a great deal: while a few genes exhibit classical induction with adriamycin, the majority of the genes are unchanged or are mildly repressed by either hypoxia or adriamycin. Further analysis using p53 mutated in the core DNA binding domain revealed that the interaction of p53 with CGIs may be occurring through both sequence-dependent and -independent mechanisms. Taken together, these experiments describe the identification of novel p53 target genes and the subsequent discovery of distinctly different expression phenomena for p53 target genes under different stress scenarios.

Original languageEnglish (US)
Pages (from-to)7030-7045
Number of pages16
JournalMolecular and Cellular Biology
Volume26
Issue number19
DOIs
StatePublished - Oct 1 2006
Externally publishedYes

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p53 Genes
Doxorubicin
Genes
HCT116 Cells
CpG Islands
Chromatin Immunoprecipitation
DNA
Proliferating Cell Nuclear Antigen
Genetic Association Studies
DNA Damage
Gene Expression
Polymerase Chain Reaction
Proteins
Hypoxia

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Functional analysis of p53 binding under differential stresses. / Krieg, Adam; Hammond, Ester M.; Giaccia, Amato J.

In: Molecular and Cellular Biology, Vol. 26, No. 19, 01.10.2006, p. 7030-7045.

Research output: Contribution to journalArticle

Krieg, Adam ; Hammond, Ester M. ; Giaccia, Amato J. / Functional analysis of p53 binding under differential stresses. In: Molecular and Cellular Biology. 2006 ; Vol. 26, No. 19. pp. 7030-7045.
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