Identification of NCF2/p67phox as a novel p53 target gene

Dafne Italiano, Anna Maria Lena, Gerry Melino, Eleonora Candi

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Analysis of microarrays performed in p53-, TAp63α- and ΔNp63α-inducible SaOs-2 cell lines allowed the identification of NCF2 mRNA upregulation in response to p53 induction. NCF2 gene encodes for p67phox, the cytosolic subunit of the NADPH oxidase enzyme complex. The recruitment of p67phox to the cell membrane causes the activation of the NADPH oxidase complex followed by the generation of NADP+ and superoxide from molecular oxygen. The presence of three putative p53 binding sites on the NCF2 promoter was predicted, and the subsequent luciferase and chromatin immunoprecipitation assays showed the activation of NCF2 promoter by p53 and its direct binding in vivo to at least one of the sites, thus confirming the hypothesis. NCF2 upregulation was also confirmed by real-time PCR in several cell lines after p53 activation. NCF2 knockdown by siRNA results in a significant reduction of ROS production and stimulates cell death, suggesting a protective function of Nox2-generated ROS in cells against apoptosis. These results provide insight into the redox-sensitive signaling mechanism that mediates cell survival involving p53 and its novel target NCF2/p67phox.

Original languageEnglish
Pages (from-to)4589-4596
Number of pages8
JournalCell Cycle
Volume11
Issue number24
DOIs
Publication statusPublished - Dec 15 2012

Fingerprint

p53 Genes
NADPH Oxidase
Up-Regulation
Cell Line
Chromatin Immunoprecipitation
Microarray Analysis
Luciferases
NADP
Superoxides
Small Interfering RNA
Oxidation-Reduction
Real-Time Polymerase Chain Reaction
Cell Survival
Cell Death
Binding Sites
Cell Membrane
Apoptosis
Oxygen
Messenger RNA
Enzymes

Keywords

  • Apoptosis
  • NADPH oxidase
  • p53
  • p53 family members
  • Reactive oxygen species

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Developmental Biology

Cite this

Identification of NCF2/p67phox as a novel p53 target gene. / Italiano, Dafne; Lena, Anna Maria; Melino, Gerry; Candi, Eleonora.

In: Cell Cycle, Vol. 11, No. 24, 15.12.2012, p. 4589-4596.

Research output: Contribution to journalArticle

Italiano, Dafne ; Lena, Anna Maria ; Melino, Gerry ; Candi, Eleonora. / Identification of NCF2/p67phox as a novel p53 target gene. In: Cell Cycle. 2012 ; Vol. 11, No. 24. pp. 4589-4596.
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