Publication details
Recognition of non-canonical DNA structures in genomic DNA sequences by p53 proteins
| Authors | |
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| Year of publication | 2009 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | In our analyses we combined molecular and computational approaches to understand the mutant p53 function in specific gene regulation via binding to non-canonical DNA structures in chromatin DNA. We used two glioblastoma cell lines U251 (R273H) and Onda11 (R273C) expressing endogenous mutp53 proteins to isolate natural mutant p53 binding sites (mutp53BS) by genome- wide ChIP-cloning. In our computational work, we developed tools for rapid identification of DNA sequences (among the isolated mutp53BS) tending to form non-B structures (hairpins and triplex DNA) as well as for mapping their genomic locations. These sites are frequently localized in the regulatory first introns of genes and are enriched in repetitive elements. Potential to form triplex and cruciform structures was predicted by developed computational tools and detected by enzymatic and chemical probing. The role of topological status of studied DNA and p53 domains in mutp53BS recognition was investigated by recombinant mutp53 proteins in vitro and reporter assays in vivo. Our data suggest that the mutant p53 proteins bind selectively non-B DNA structures not only in vitro but also with functional consequences in vivo. |
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