Patterns of Proteins that Associate with p53 or with p53 Binding Sites Present in the Ribosomal Gene Cluster and MDM2 (P2) Promoter
Abstract
It has been shown that p53 binding to the MDM2P2 site changes the footprint of an existing nearby TA box In addition, it has been shown that p53 has the ability to associate with other proteins. By DNA affi ity chromatography% we studied the binding of p53 to two p53-binding sites. One of them is present in the Promoter 2 of the MDM2 gene and the other one is an ideal p53-binding site known as superconsensus sequence (SCS). Our results showed that p53 can be purified by DNA affinity chromatography using eit r site. We also showed that, the TATA Binding Protein (TBP) and the TBP associated factors TAFII40 an TAFII6O coeluted with p53 from the DNA affinity columns. The fractions from the MDM2 column sho ed an activity able to supershift the TBP complex when analyzed by gel shift using a TATA box as a probe. We could not detect this supershift when p53 was eluted from the SCS column. Only a fraction of the p53 loaded was bound, suggesting that the DNA Sites select for p53 subpopulations. To the best of our knowledge, this is the first time that p53-associated proteins are being studied by DNA affinity chromatography.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 2000
- Accession Number
- ADB267156
Entities
People
- Maria J. Molina
Organizations
- Hunter College