Abstract
Several promoters have been shown to have sequence specifically positioned nucleosomes that determine the architecture of the promoter. DNA binding proteins that regulate gene expression are in many cases known to bind to their cognate DNA segments organized within such positioned nucleosomes. It has become increasingly evident that the cooperation of chromatin and transcription factors results in an efficient and fine-tuned regulation of transcription. The first step in a gene induction event must be the access of transcription factors for the regulatory promoter/enhancer target sites. In this perspective it becomes interesting to evaluate the affinity of DNA binding proteins for their cognate binding site in a nucleosome context. Here we describe the preparation of nucleosome probe, a method for in vitro nucleosome reconstitution by salt dilution, purification of the reconstituted mononucleosomes, and characterization of the translational and rotational positions of the nucleosomal DNA. In addition, methods for affinity determination and characterization of protein-nucleosomal DNA interaction, such as methylation protection and methylation interference by dimethyl sulfate, quantitative DNase I footprinting, and electrophoreti...Continue Reading
References
Feb 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·A M Maxam, W Gilbert
Sep 15, 1978·Journal of Molecular Biology·L C Lutter
May 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·T Perlmann
Aug 1, 1991·Journal of Structural Biology·P ErikssonO Wrange
Feb 1, 1991·Molecular and Cellular Biology·T K ArcherG L Hager
Jul 1, 1991·Genes & Development·I C TaylorR E Kingston
Mar 9, 1990·Cell·B PiñaM Beato
May 1, 1990·Electrophoresis·R F JohnstonD L Barker
Oct 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·T E Shrader, D M Crothers
Oct 21, 1988·Cell·S Y TsaiB W O'Malley
Apr 24, 1987·Cell·Y LorchR D Kornberg
Aug 28, 1987·Cell·R Losa, D D Brown
Jan 1, 1986·Methods in Enzymology·M BrenowitzG K Ackers
Dec 5, 1986·Cell·K GorskiU Schibler
Aug 1, 1984·Cell·K S Zaret, K R Yamamoto
May 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·C Scheidereit, M Beato
Aug 1, 1995·Molecular and Cellular Biology·Q Li, O Wrange
Dec 1, 1993·Genes & Development·Q Li, O Wrange
Jan 15, 1993·Cell·D Y LeeA P Wolffe
Nov 25, 1995·Nucleic Acids Research·J S GoddeA P Wolffe
Nov 1, 1974·Nucleic Acids Research·M Noll
Citations
Aug 19, 2007·Nature Protocols·Lance M Hellman, Michael G Fried
Feb 28, 2001·The EMBO Journal·M Ouhammouch, E P Geiduschek
Jan 26, 2002·Nucleic Acids Research·Isabell Dahlke, Michael Thomm
Aug 13, 2011·Cell·Ty C VossGordon L Hager
Feb 3, 2009·Molecular Immunology·Natalia PetrakovaMaria Isaguliants
Apr 25, 2001·The Journal of Biological Chemistry·M HäkliJ J Palvimo
Aug 8, 1998·The Journal of Biological Chemistry·C Thiriet, J J Hayes
May 16, 2020·Trends in Cell Biology·David M Suter