title: Differential salt-induced dissociation of the p53 protein complexes with circular and linear plasmid DNA substrates suggest involvement of a sliding mechanism.
abstract: A study of the effects of salt conditions on the association and dissociation of wild type p53 with different ~3 kbp long plasmid DNA substrates (supercoiled, relaxed circular and linear, containing or lacking a specific p53 binding site, p53CON) using immunoprecipitation at magnetic beads is presented. Salt concentrations above 200 mM strongly affected association of the p53 protein to any plasmid DNA substrate. Strikingly different behavior was observed when dissociation of pre-formed p53-DNA complexes in increased salt concentrations was studied. While contribution from the p53CON to the stability of the p53-DNA complexes was detected between 100 and 170 mM KCl, p53 complexes with circular DNAs (but not linear) exhibited considerable resistance towards salt treatment for KCl concentrations as high as 2 M provided that the p53 basic C-terminal DNA binding site (CTDBS) was available for DNA binding. On the contrary, when the CTDBS was blocked by antibody used for immunoprecipitation, all p53-DNA complexes were completely dissociated from the p53 protein in KCl concentrations≥200 mM under the same conditions. These observations suggest: (a) different ways for association and dissociation of the p53-DNA complexes in the presence of the CTDBS; and (b) a critical role for a sliding mechanism, mediated by the C-terminal domain, in the dissociation process.
authors: Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i., Královopolská 135, Brno CZ-612 65, Czech Republic. besta@ibp.cz.
authors: Peter
authors: P
authors: Šebest
authors: Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i., Královopolská 135, Brno CZ-612 65, Czech Republic. maruska@ipb.cz.
authors: Marie
authors: M
authors: Brázdová
authors: Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i., Královopolská 135, Brno CZ-612 65, Czech Republic. miroslav.fojta@ceitec.muni.cz.|Central European Institute of Technology, Masaryk University, Kamenice 753/5, Brno CZ-625 00, Czech Republic. miroslav.fojta@ceitec.muni.cz.
authors: Miroslav
authors: M
authors: Fojta
authors: Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i., Královopolská 135, Brno CZ-612 65, Czech Republic. hapi@ibp.cz.
authors: Hana
authors: H
authors: Pivoňková
mesh_terms: D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism*; D011189:Potassium Chloride / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D012492:Salts / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: D012492:Salts; D016159:Tumor Suppressor Protein p53; D011189:Potassium Chloride
references: Nat Rev Cancer. 2009 Oct;9(10):714-23
references: 19730431
references: Nat Rev Cancer. 2009 Oct;9(10):724-37
references: 19776742
references: Biochem Biophys Res Commun. 2010 Jan 15;391(3):1409-14
references: 20026061
references: Biochem Biophys Res Commun. 2010 Mar 19;393(4):894-9
references: 20175992
references: Anal Chim Acta. 2010 Jun 4;668(2):166-70
references: 20493293
references: Proc Natl Acad Sci U S A. 2011 Jan 11;108(2):563-8
references: 21178072
references: Proc Natl Acad Sci U S A. 2011 May 17;108(20):8251-6
references: 21525412
references: EMBO J. 2011 Jun 1;30(11):2167-76
references: 21522129
references: J Am Chem Soc. 2012 Sep 5;134(35):14555-62
references: 22880817
references: Proc Natl Acad Sci U S A. 2012 Oct 9;109(41):16552-7
references: 23012405
references: PLoS One. 2013;8(3):e59567
references: 23555710
references: Anal Biochem. 2013 Nov 1;442(1):51-61
references: 23928048
references: Biochem Biophys Res Commun. 2013 Nov 8;441(1):83-8
references: 24134839
references: Anal Bioanal Chem. 2014 Sep;406(24):5843-52
references: 25056872
references: Nucleic Acids Res. 2014 Nov 10;42(20):12404-14
references: 25324308
references: Eur J Biochem. 2001 Feb;268(3):573-81
references: 11168396
references: Biochim Biophys Acta. 2001 Feb 16;1517(3):392-7
references: 11342217
references: Nucleic Acids Res. 2002 Nov 15;30(22):4966-74
references: 12434001
references: Oncogene. 2004 Mar 18;23(12):2119-27
references: 14755248
references: J Mol Biol. 2004 Aug 27;341(5):1145-59
references: 15321712
references: Eur J Biochem. 2004 Oct;271(19):3865-76
references: 15373832
references: J Immunol Methods. 1992 Jul 6;151(1-2):237-44
references: 1378473
references: Methods Enzymol. 1992;212:105-20
references: 1518443
references: Nat Genet. 1992 Apr;1(1):45-9
references: 1301998
references: Biochem J. 1994 Mar 15;298 Pt 3:513-6
references: 8141761
references: Oncogene. 1997 Aug 14;15(7):857-69
references: 9266973
references: Oncogene. 1997 Oct;15(18):2201-9
references: 9393978
references: Mol Cell. 2004 Nov 5;16(3):413-24
references: 15525514
references: Nucleic Acids Res. 2005;33(3):1087-100
references: 15722483
references: J Mol Biol. 2005 May 6;348(3):641-53
references: 15826661
references: Proc Natl Acad Sci U S A. 2005 Nov 1;102(44):15883-8
references: 16243975
references: Biochem Biophys Res Commun. 2006 Jan 13;339(2):477-84
references: 16300733
references: Biochem Biophys Res Commun. 2006 Mar 10;341(2):470-7
references: 16426567
references: Oncogene. 2007 Apr 2;26(15):2226-42
references: 17401432
references: Biochem J. 2008 May 15;412(1):57-63
references: 18271758
references: Biophys J. 2008 Jul;95(1):L01-3
references: 18424488
references: Anal Chem. 2008 Oct 1;80(19):7609-13
references: 18778033