Cited by 360Cancer-related Molecular PathwaysCell death mechanisms and regulationGenomics and Chromatin Dynamics

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Wild-type p53 transactivates the KILLER/DR5 gene through an intronic sequence-specific DNA-binding site

Takimoto, R, El-Deiry, W S·2000·Oncogene

DOI 10.1038/sj.onc.1203489

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Abstract

KILLER/DR5, a tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor gene, has been shown to be induced by DNA damaging agents and radiation in a p53-dependent manner. Although TRAIL is a potential therapeutic agent for cancer, the induction mechanism of its receptors is poorly understood. Here we show the identification of three p53 DNA-binding sites in the KILLER/DR5 genomic locus located upstream (BS1; -0.82 Kb) of the ATG site, within Intron 1 (BS2; +0.25 Kb downstream of the ATG) and within Intron 2 (BS3; +1.25 Kb downstream of the ATG). A modified p53-binding and immunoselection protocol using a wild-type p53-expressing adenovirus vector (Ad-p53) was used to identify the binding sites and to show that each binding site can bind specifically to wild-type p53 protein (wt-p53). A reporter assay revealed that only BS2 could enhance luciferase expression driven by a basal promoter. We constructed a reporter plasmid carrying the genomic regulatory region of KILLER/DR5 including the three p53 DNA-binding sites but no additional basal promoter. The genomic fragment showed basal transcriptional activity which was induced by wt-p53 but not by mutant p53, and human papilloma virus E6 inhibited the p53-dependent activation. Mutation of BS2 abrogated not only the binding activity of wt-p53 but also the induction of the KILLER/DR5 genomic promoter-reporter gene, indicating that BS2 is responsible for the p53-dependent transactivation of KILLER/ DR5. In p53-wild-type but not -mutant or -null cell lines, doxorubicin treatment stabilized p53 protein, and increased specific binding to BS2 as revealed by EMSA, and upregulated the KILLER/DR5 promoter-luciferase reporter gene. These results suggest that the transactivation of KILLER/DR5 is directly regulated by exogenous or endogenous wt-p53 and establishes KILLER/DR5 as a p53 target gene that can signal apoptotic death.

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Self-citation

Authors: Takimoto, R., & El-Deiry, W. S.
Affiliations: Howard Hughes Medical Institute (US)
Venue: Oncogene
Publisher: Springer Science and Business Media LLC
Published: 2000-03-30
Volume: 19
Issue: 14
Pages: 1735-1743
ISSN: 0950-9232 (ISSN-L)
1476-5594
DOI: 10.1038/sj.onc.1203489
OpenAlex: W2053301688
Cited by: 360
References: 39
Citations by year: 2026: 3 · 2025: 6 · 2024: 3 · 2023: 12 · 2022: 5 · 2021: 6 · 2020: 11 · 2019: 10 · 2018: 7 · 2017: 10 · 2016: 8 · 2015: 10 · 2014: 12 · 2013: 8 · 2012: 18
FWCI: 7.78
Type: journal article
Language: en
Open access: closed
Licence: www.springer.com/tdm (tdm)
www.springer.com/tdm (vor)
Locations: Oncogene (publishedVersion)
PubMed (publishedVersion)
Funders: Howard Hughes Medical Institute
Japan Health Sciences Foundation
National Cancer Institute (P01CA075138, P01 CA75138-01)
Topics: Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Keywords: p53, DR5, TRAIL, p53 target genes, intronic enhancer, transcriptional activation, doxorubicin, p53 stabilization, apoptosis, E6 oncoprotein
MeSH terms: Binding Sites, DNA, DNA Damage, Doxorubicin, Humans, Introns, Oncogene Proteins, Viral, Promoter Regions, Genetic, Tumor Cells, Cultured, Transcriptional Activation, Gene Expression, Tumor Suppressor Protein p53, Mutagenesis, Apoptosis, Genes, Reporter, Receptors, Tumor Necrosis Factor, Receptors, TNF-Related Apoptosis-Inducing Ligand

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Wild-type p53 transactivates the KILLER/DR5 gene through an intronic sequence-specific DNA-binding site

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