Three assays show differences in binding of wild-type and mutant p53 to unique gene sequences

Uma Chandrachud, Susannah Gal

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Cancer-associated mutations in the p53 gene often change amino acids in the protein's DNA binding domain. We used three different binding assays specifically gel shift, DNA binding scintillation proximity assay and a streptavidin magnetic bead assay to analyze the DNA binding of the tumor suppressor p53 from 4 human cell lines with different DNA sequences from the mdm2, p21 and cyclin G genes and a mutant form of the cyclin G sequence. Treatment of MCF-7 cells having wild-type p53 with hydrogen peroxide increased the binding of p53 to DNA as detected using all three assays, but to different extents. The p53 proteins from the thyroid cancer cell lines with different p53 mutations (ARO, WRO and NPA) have comparable binding reactions in the three assays, but show different specificities for the sequences. Here we show that multiple different binding assays allow us to generate a more complete picture of the function of DNA transcription factors in diseases such as cancer.

Original languageEnglish (US)
Pages (from-to)445-454
Number of pages10
JournalTechnology in Cancer Research and Treatment
Volume8
Issue number6
StatePublished - Dec 1 2009

Fingerprint

Cyclin G
DNA
Genes
Cell Line
Neoplasms
Mutation
Streptavidin
p53 Genes
MCF-7 Cells
DNA-Binding Proteins
Thyroid Neoplasms
Hydrogen Peroxide
Transcription Factors
Gels
Amino Acids
Proteins

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

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Three assays show differences in binding of wild-type and mutant p53 to unique gene sequences. / Chandrachud, Uma; Gal, Susannah.

In: Technology in Cancer Research and Treatment, Vol. 8, No. 6, 01.12.2009, p. 445-454.

Research output: Contribution to journalArticle

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