EurekaMag.com logo
+ Site Statistics
References:
52,725,316
Abstracts:
28,411,598
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

Preferential binding of hot spot mutant p53 proteins to supercoiled DNA in vitro and in cells






Plos One 8(3): E59567-E59567

Preferential binding of hot spot mutant p53 proteins to supercoiled DNA in vitro and in cells

Hot spot mutant p53 (mutp53) proteins exert oncogenic gain-of-function activities. Binding of mutp53 to DNA is assumed to be involved in mutp53-mediated repression or activation of several mutp53 target genes. To investigate the importance of DNA topology on mutp53-DNA recognition in vitro and in cells, we analyzed the interaction of seven hot spot mutp53 proteins with topologically different DNA substrates (supercoiled, linear and relaxed) containing and/or lacking mutp53 binding sites (mutp53BS) using a variety of electrophoresis and immunoprecipitation based techniques. All seven hot spot mutp53 proteins (R175H, G245S, R248W, R249S, R273C, R273H and R282W) were found to have retained the ability of wild-type p53 to preferentially bind circular DNA at native negative superhelix density, while linear or relaxed circular DNA was a poor substrate. The preference of mutp53 proteins for supercoiled DNA (supercoil-selective binding) was further substantiated by competition experiments with linear DNA or relaxed DNA in vitro and ex vivo. Using chromatin immunoprecipitation, the preferential binding of mutp53 to a sc mutp53BS was detected also in cells. Furthermore, we have shown by luciferase reporter assay that the DNA topology influences p53 regulation of BAX and MSP/MST1 promoters. Possible modes of mutp53 binding to topologically constrained DNA substrates and their biological consequences are discussed.


Accession: 055116778

PMID: 23555710

DOI: 10.1371/journal.pone.0059567



Related references

Preferential binding of bacteriophage Mu repressor to supercoiled Mu DNA. Plasmid 13(3): 173-181, 1985

Assays for the preferential binding of human topoisomerase I to supercoiled DNA. Methods in Molecular Biology 582: 49-57, 2010

Mutational analysis of the preferential binding of human topoisomerase I to supercoiled DNA. Febs Journal 276(20): 5906-5919, 2009

Preferential binding of the archaebacterial histone-like MC1 protein to negatively supercoiled DNA minicircles. Biochemistry 35(24): 7954-7958, 1996

Preferential binding of E.coli histone-like protein HU alpha to negatively supercoiled DNA. Nucleic Acids Research 20(7): 1553-1558, 1992

Preferential binding of escherichia coli histone like protein hu alpha to negatively supercoiled dna. Nucleic Acids Research 20(7): 1553-1558, 1992

Preferential binding of tumor suppressor p53 to positively or negatively supercoiled DNA involves the C-terminal domain. Journal of Molecular Biology 292(2): 241-249, 1999

Lipopolysaccharide (LPS) binding to 73-kDa and 38-kDa surface proteins on lymphoreticular cells: Preferential inhibition of LPS binding to the former by Rhodopseudomonas sphaeroides lipid A. Immunology Letters 36(3): 245-250, 1993

Mutant p53 proteins bind selectively supercoiled DNA. 2007

Distinct Rayleigh scattering from hot spot mutant p53 proteins reveals cancer cells. Small 10(14): 2954-2962, 2015