Journal
TECHNOLOGY IN CANCER RESEARCH & TREATMENT
Volume 8, Issue 6, Pages 445-453Publisher
SAGE PUBLICATIONS INC
DOI: 10.1177/153303460900800606
Keywords
DNA affinity immunoblotting; DNA binding; Electrophoretic mobility shift assay (EMSA); Scintillation proximity assay (SPA) beads; Streptavidin magnetic beads
Categories
Funding
- NIH [1R15 CA101783-01A1]
- US Department of Energy [DE-FG02-06ER64281]
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available